Modelling the zebrafish gut microbiome’s resistance and sensitivity to climate change and infection

Main Figures and Supplemental Results

Overview

Publication Information

The published version of this manuscript can be found here: Sieler Jr M.J., Al-Samarrie C.E., Kasschau K.D., Kent M.L., Sharpton T.J.. Modeling the zebrafish gut microbiome’s resistance and sensitivity to climate change and infection. bioRxiv (2025). DOI: https://doi.org/10.1101/2025.03.28.644597

All code generated during this analysis is available in the GitHub Repository.

The raw sequence files generated during the current study are available at the NCBI Sequence Read Archive (SRA) project numbers: PRJNA1219243.

Fig. 1) Experimental Design Schematic

Fig.1 Experimental design showing treatments and husbandry events during the course of the study. Symbols indicate when an experimental event occurred at each time point. (1) 260 fish were assigned and acclimated to one of three water temperature groups (e.g., 28°C, 32°C, or 35°C) and reared from 0 to 164 days post fertilization (dpf). (2a) At 164 dpf (or 0 days post exposure; DPE), fecal collections were collected from a random selection of five fish per tank (n = 60). Additionally, histological and wet mount assessments were conducted on selected fish to assess presence of infection and infection burden. (1b) Afterwards, a cohort of fish from each water temperature group were exposed to Pseudocapillaria tomentosa. (3-6) Subsequent fecal samples were collected and histopathological assessments were conducted at 14 dpe (n = 54), (3) 21 dpe (n = 48), (4) 28 dpe (n = 47), and (5) 42 dpe (n = 51).

Results

Fig. 2) Water temperature shapes gut microbiome structure

Fig. 2 Effects of water temperature on zebrafish gut microbiomes. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main effect of temperature. The analysis shows that gut microbiome composition significantly differs between fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity increases with time from 0 days post exposure (dpe) to 42 dpe, irrespective of water temperature. (D) Capscale ordination of gut microbiome composition based on the Bray-Curtis dissimilarity constrained on the main effects of water temperature and time (days post exposure, dpe), and their interaction. The analysis shows that shows that gut microbiome composition differs between fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated by covariates.

2A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature); Unexposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.754	0.091	−8.309	<0.001	****
Temperature32	0.051	0.127	0.400	≥0.25	ns
Temperature35	0.054	0.129	0.421	≥0.25	ns
Simpson
(Intercept)	−0.101	0.092	−1.103	≥0.25	ns
Temperature32	0.132	0.128	1.030	≥0.25	ns
Temperature35	0.340	0.131	2.591	0.010	*
Richness
(Intercept)	−0.231	0.075	−3.079	0.002	**
Temperature32	0.047	0.105	0.446	≥0.25	ns
Temperature35	−0.473	0.110	−4.299	<0.001	****
Phylogenetic
(Intercept)	−0.255	0.072	−3.540	<0.001	***
Temperature32	0.068	0.101	0.673	≥0.25	ns
Temperature35	−0.475	0.106	−4.482	<0.001	****

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Unexposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	0.225	2.000	≥0.25	ns
Simpson
Temperature	6.830	2.000	0.033	*
Richness
Temperature	27.512	2.000	<0.001	****
Phylogenetic
Temperature	31.449	2.000	<0.001	****

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Unexposed fish
term	.y.	group1	group2	estimate	std.error	statistic	adj.p.value	Variable
Shannon
Temperature	Alpha.Score	32	28	0.051	0.127	0.400	≥0.25	Temperature
Temperature	Alpha.Score	35	28	0.054	0.129	0.421	≥0.25	Temperature
Temperature	Alpha.Score	35	32	0.004	0.127	0.029	≥0.25	Temperature
Simpson
Temperature	Alpha.Score	32	28	0.132	0.128	1.030	≥0.25	Temperature
Temperature	Alpha.Score	35	28	0.340	0.131	2.591	0.026	Temperature
Temperature	Alpha.Score	35	32	0.207	0.130	1.596	0.247	Temperature
Richness
Temperature	Alpha.Score	32	28	0.047	0.105	0.446	≥0.25	Temperature
Temperature	Alpha.Score	35	28	−0.473	0.110	−4.299	<0.001	Temperature
Temperature	Alpha.Score	35	32	−0.520	0.109	−4.768	<0.001	Temperature
Phylogenetic
Temperature	Alpha.Score	32	28	0.068	0.101	0.673	≥0.25	Temperature
Temperature	Alpha.Score	35	28	−0.475	0.106	−4.482	<0.001	Temperature
Temperature	Alpha.Score	35	32	−0.543	0.105	−5.172	<0.001	Temperature

Addl. Metrics

2B)

Tables

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature); Unexposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	2.294	0.133	12.900	0.001	***
Residual	168.000	14.936	0.867	NA	NA	NA
Total	170.000	17.229	1.000	NA	NA	NA
canberra
Temperature	2.000	3.862	0.099	9.178	0.001	***
Residual	168.000	35.344	0.901	NA	NA	NA
Total	170.000	39.206	1.000	NA	NA	NA
gunifrac
Temperature	2.000	1.422	0.102	9.566	0.001	***
Residual	168.000	12.485	0.898	NA	NA	NA
Total	170.000	13.907	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temperature	2.000	0.008	0.004	0.386	≥0.25	ns
Residual	168.000	1.771	0.011	NA	NA	NA
canberra
Temperature	2.000	0.008	0.004	0.873	≥0.25	ns
Residual	168.000	0.759	0.005	NA	NA	NA
gunifrac
Temperature	2.000	0.005	0.003	0.369	≥0.25	ns
Residual	168.000	1.143	0.007	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Unexposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temperature	32	28	−0.011	−0.056	0.034	≥0.25	ns
Distance	Temperature	35	28	0.005	−0.041	0.051	≥0.25	ns
Distance	Temperature	35	32	0.016	−0.029	0.062	≥0.25	ns
canberra
Distance	Temperature	32	28	−0.001	−0.031	0.028	≥0.25	ns
Distance	Temperature	35	28	0.014	−0.016	0.044	≥0.25	ns
Distance	Temperature	35	32	0.015	−0.015	0.045	≥0.25	ns
gunifrac
Distance	Temperature	32	28	−0.013	−0.049	0.023	≥0.25	ns
Distance	Temperature	35	28	−0.004	−0.040	0.033	≥0.25	ns
Distance	Temperature	35	32	0.009	−0.027	0.046	≥0.25	ns

Addl. Metrics

2C)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*DPE); Unexposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−1.074	0.138	−7.754	<0.001	****
Temperature32	0.075	0.193	0.386	≥0.25	ns
Temperature35	0.154	0.193	0.797	≥0.25	ns
DPE	0.018	0.006	3.086	0.002	**
Temperature32:DPE	−0.001	0.008	−0.170	≥0.25	ns
Temperature35:DPE	−0.005	0.008	−0.585	≥0.25	ns
Simpson
(Intercept)	−0.433	0.134	−3.245	0.001	**
Temperature32	0.157	0.186	0.841	≥0.25	ns
Temperature35	0.408	0.187	2.177	0.031	*
DPE	0.019	0.006	3.293	0.001	**
Temperature32:DPE	−0.001	0.008	−0.173	≥0.25	ns
Temperature35:DPE	−0.003	0.008	−0.343	≥0.25	ns
Richness
(Intercept)	−0.651	0.108	−6.017	<0.001	****
Temperature32	0.189	0.150	1.257	0.211	ns
Temperature35	−0.059	0.154	−0.383	≥0.25	ns
DPE	0.024	0.005	5.137	<0.001	****
Temperature32:DPE	−0.008	0.006	−1.251	0.213	ns
Temperature35:DPE	−0.024	0.007	−3.476	<0.001	***
Phylogenetic
(Intercept)	−0.709	0.102	−6.970	<0.001	****
Temperature32	0.237	0.141	1.682	0.095	ns
Temperature35	0.010	0.144	0.072	≥0.25	ns
DPE	0.026	0.004	5.907	<0.001	****
Temperature32:DPE	−0.010	0.006	−1.579	0.116	ns
Temperature35:DPE	−0.028	0.006	−4.344	<0.001	****

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Unexposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	0.322	2.000	≥0.25	ns
DPE	22.895	1.000	<0.001	****
Temperature:DPE	0.363	2.000	≥0.25	ns
Simpson
Temperature	8.551	2.000	0.014	*
DPE	28.161	1.000	<0.001	****
Temperature:DPE	0.118	2.000	≥0.25	ns
Richness
Temperature	31.759	2.000	<0.001	****
DPE	27.201	1.000	<0.001	****
Temperature:DPE	12.460	2.000	0.002	**
Phylogenetic
Temperature	38.358	2.000	<0.001	****
DPE	31.813	1.000	<0.001	****
Temperature:DPE	19.500	2.000	<0.001	****

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Unexposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	DPE	14	0	1.065	0.294	3.625	0.003	DPE	28
28	Alpha.Score	DPE	21	0	0.722	0.281	2.566	0.076	DPE	28
28	Alpha.Score	DPE	28	0	1.187	0.274	4.335	<0.001	DPE	28
28	Alpha.Score	DPE	42	0	0.548	0.296	1.853	≥0.25	DPE	28
28	Alpha.Score	DPE	21	14	−0.343	0.317	−1.083	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.122	0.310	0.392	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	−0.517	0.330	−1.567	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	0.465	0.299	1.558	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	−0.174	0.319	−0.545	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	−0.639	0.312	−2.045	0.243	DPE	28
32	Alpha.Score	DPE	14	0	0.006	0.250	0.022	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	0.112	0.247	0.452	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.420	0.248	1.695	≥0.25	DPE	32
32	Alpha.Score	DPE	42	0	0.694	0.236	2.943	0.026	DPE	32
32	Alpha.Score	DPE	21	14	0.106	0.286	0.371	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.414	0.287	1.445	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.688	0.276	2.490	0.092	DPE	32
32	Alpha.Score	DPE	28	21	0.308	0.283	1.087	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.582	0.273	2.131	0.205	DPE	32
32	Alpha.Score	DPE	42	28	0.274	0.274	1.001	≥0.25	DPE	32
35	Alpha.Score	DPE	14	0	−0.006	0.210	−0.027	≥0.25	DPE	35
35	Alpha.Score	DPE	21	0	−0.249	0.218	−1.145	≥0.25	DPE	35
35	Alpha.Score	DPE	28	0	0.232	0.204	1.138	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	0.661	0.207	3.194	0.012	DPE	35
35	Alpha.Score	DPE	21	14	−0.243	0.253	−0.962	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.238	0.242	0.985	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.667	0.244	2.733	0.049	DPE	35
35	Alpha.Score	DPE	28	21	0.481	0.248	1.938	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.910	0.251	3.628	0.003	DPE	35
35	Alpha.Score	DPE	42	28	0.429	0.239	1.792	≥0.25	DPE	35
Simpson
28	Alpha.Score	DPE	14	0	1.256	0.273	4.611	<0.001	DPE	28
28	Alpha.Score	DPE	21	0	0.761	0.247	3.076	0.018	DPE	28
28	Alpha.Score	DPE	28	0	1.334	0.254	5.241	<0.001	DPE	28
28	Alpha.Score	DPE	42	0	0.514	0.257	2.002	≥0.25	DPE	28
28	Alpha.Score	DPE	21	14	−0.496	0.302	−1.644	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.077	0.308	0.251	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	−0.742	0.310	−2.398	0.114	DPE	28
28	Alpha.Score	DPE	28	21	0.573	0.285	2.008	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	−0.246	0.288	−0.857	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	−0.819	0.294	−2.789	0.042	DPE	28
32	Alpha.Score	DPE	14	0	0.065	0.249	0.261	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	0.142	0.249	0.569	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.592	0.261	2.272	0.152	DPE	32
32	Alpha.Score	DPE	42	0	0.724	0.254	2.850	0.035	DPE	32
32	Alpha.Score	DPE	21	14	0.077	0.287	0.267	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.527	0.297	1.773	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.659	0.292	2.260	0.156	DPE	32
32	Alpha.Score	DPE	28	21	0.451	0.297	1.517	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.582	0.291	1.999	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	0.132	0.302	0.436	≥0.25	DPE	32
35	Alpha.Score	DPE	14	0	0.149	0.219	0.683	≥0.25	DPE	35
35	Alpha.Score	DPE	21	0	−0.167	0.218	−0.766	≥0.25	DPE	35
35	Alpha.Score	DPE	28	0	0.413	0.222	1.858	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	0.836	0.244	3.431	0.005	DPE	35
35	Alpha.Score	DPE	21	14	−0.316	0.257	−1.232	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.263	0.260	1.013	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.687	0.279	2.465	0.097	DPE	35
35	Alpha.Score	DPE	28	21	0.579	0.260	2.233	0.165	DPE	35
35	Alpha.Score	DPE	42	21	1.003	0.278	3.604	0.003	DPE	35
35	Alpha.Score	DPE	42	28	0.424	0.282	1.506	≥0.25	DPE	35
Richness
28	Alpha.Score	DPE	14	0	0.432	0.236	1.836	≥0.25	DPE	28
28	Alpha.Score	DPE	21	0	0.550	0.218	2.526	0.084	DPE	28
28	Alpha.Score	DPE	28	0	0.767	0.218	3.525	0.004	DPE	28
28	Alpha.Score	DPE	42	0	0.970	0.227	4.278	<0.001	DPE	28
28	Alpha.Score	DPE	21	14	0.118	0.263	0.449	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.335	0.262	1.275	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	0.537	0.270	1.990	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	0.217	0.247	0.880	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	0.419	0.255	1.647	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	0.203	0.254	0.796	≥0.25	DPE	28
32	Alpha.Score	DPE	14	0	−0.170	0.177	−0.965	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	−0.026	0.175	−0.147	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.187	0.180	1.041	≥0.25	DPE	32
32	Alpha.Score	DPE	42	0	0.769	0.176	4.359	<0.001	DPE	32
32	Alpha.Score	DPE	21	14	0.145	0.204	0.711	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.358	0.208	1.721	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.939	0.205	4.585	<0.001	DPE	32
32	Alpha.Score	DPE	28	21	0.213	0.206	1.031	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.794	0.203	3.906	<0.001	DPE	32
32	Alpha.Score	DPE	42	28	0.581	0.208	2.801	0.040	DPE	32
35	Alpha.Score	DPE	14	0	−0.560	0.214	−2.612	0.067	DPE	35
35	Alpha.Score	DPE	21	0	−0.551	0.214	−2.578	0.073	DPE	35
35	Alpha.Score	DPE	28	0	0.110	0.196	0.561	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	0.007	0.206	0.036	≥0.25	DPE	35
35	Alpha.Score	DPE	21	14	0.008	0.259	0.032	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.670	0.245	2.733	0.048	DPE	35
35	Alpha.Score	DPE	42	14	0.567	0.253	2.240	0.162	DPE	35
35	Alpha.Score	DPE	28	21	0.661	0.245	2.702	0.052	DPE	35
35	Alpha.Score	DPE	42	21	0.559	0.253	2.210	0.173	DPE	35
35	Alpha.Score	DPE	42	28	−0.103	0.238	−0.431	≥0.25	DPE	35
Phylogenetic
28	Alpha.Score	DPE	14	0	0.452	0.229	1.969	≥0.25	DPE	28
28	Alpha.Score	DPE	21	0	0.679	0.211	3.210	0.011	DPE	28
28	Alpha.Score	DPE	28	0	0.872	0.212	4.120	<0.001	DPE	28
28	Alpha.Score	DPE	42	0	1.012	0.220	4.602	<0.001	DPE	28
28	Alpha.Score	DPE	21	14	0.227	0.254	0.893	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.420	0.254	1.651	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	0.560	0.261	2.142	0.200	DPE	28
28	Alpha.Score	DPE	28	21	0.193	0.238	0.809	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	0.333	0.246	1.353	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	0.140	0.246	0.569	≥0.25	DPE	28
32	Alpha.Score	DPE	14	0	−0.030	0.164	−0.181	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	0.154	0.163	0.945	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.260	0.168	1.550	≥0.25	DPE	32
32	Alpha.Score	DPE	42	0	0.753	0.164	4.594	<0.001	DPE	32
32	Alpha.Score	DPE	21	14	0.183	0.188	0.974	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.290	0.193	1.503	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.783	0.190	4.130	<0.001	DPE	32
32	Alpha.Score	DPE	28	21	0.107	0.192	0.556	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.599	0.188	3.185	0.013	DPE	32
32	Alpha.Score	DPE	42	28	0.493	0.193	2.555	0.078	DPE	32
35	Alpha.Score	DPE	14	0	−0.393	0.204	−1.929	≥0.25	DPE	35
35	Alpha.Score	DPE	21	0	−0.453	0.206	−2.198	0.177	DPE	35
35	Alpha.Score	DPE	28	0	0.090	0.192	0.468	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	−0.111	0.204	−0.542	≥0.25	DPE	35
35	Alpha.Score	DPE	21	14	−0.059	0.246	−0.241	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.483	0.235	2.057	0.235	DPE	35
35	Alpha.Score	DPE	42	14	0.282	0.245	1.153	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	0.542	0.237	2.292	0.145	DPE	35
35	Alpha.Score	DPE	42	21	0.342	0.247	1.386	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	−0.200	0.235	−0.853	≥0.25	DPE	35

Addl. Metrics

2D)

Tables

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Unexposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	2.294	0.133	16.443	0.001	***
DPE	1.000	2.531	0.147	36.284	0.001	***
Temperature:DPE	2.000	0.896	0.052	6.425	0.001	***
Residual	165.000	11.509	0.668	NA	NA	NA
Total	170.000	17.229	1.000	NA	NA	NA
canberra
Temperature	2.000	3.862	0.099	10.667	0.001	***
DPE	1.000	3.655	0.093	20.192	0.001	***
Temperature:DPE	2.000	1.820	0.046	5.026	0.001	***
Residual	165.000	29.869	0.762	NA	NA	NA
Total	170.000	39.206	1.000	NA	NA	NA
gunifrac
Temperature	2.000	1.422	0.102	11.984	0.001	***
DPE	1.000	2.123	0.153	35.785	0.001	***
Temperature:DPE	2.000	0.574	0.041	4.840	0.001	***
Residual	165.000	9.788	0.704	NA	NA	NA
Total	170.000	13.907	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.DPE	14.000	0.290	0.021	2.348	0.006	**
Residual	156.000	1.376	0.009	NA	NA	NA
canberra
Temp.DPE	14.000	0.250	0.018	3.344	<0.001	***
Residual	156.000	0.831	0.005	NA	NA	NA
gunifrac
Temp.DPE	14.000	0.249	0.018	2.986	<0.001	***
Residual	156.000	0.929	0.006	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Unexposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.DPE	28°C__14DPE	28°C__0DPE	0.024	−0.112	0.159	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__0DPE	0.052	−0.074	0.177	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__0DPE	0.007	−0.118	0.133	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__0DPE	−0.017	−0.147	0.113	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__0DPE	0.097	−0.005	0.199	0.085	ns
Distance	Temp.DPE	32°C__14DPE	28°C__0DPE	0.041	−0.085	0.166	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__0DPE	−0.018	−0.143	0.108	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__0DPE	0.028	−0.102	0.158	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__0DPE	0.017	−0.108	0.142	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__0DPE	0.044	−0.058	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__0DPE	0.004	−0.126	0.134	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__0DPE	−0.004	−0.134	0.126	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__0DPE	0.033	−0.097	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__0DPE	0.146	0.011	0.282	0.021	*
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	0.028	−0.126	0.181	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.016	−0.170	0.137	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.040	−0.198	0.117	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__14DPE	0.073	−0.062	0.209	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	0.017	−0.136	0.171	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.041	−0.195	0.112	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	0.005	−0.153	0.162	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.006	−0.160	0.147	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__14DPE	0.020	−0.115	0.156	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.019	−0.176	0.138	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.028	−0.185	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	0.009	−0.148	0.166	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	0.123	−0.039	0.284	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.044	−0.189	0.101	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	−0.068	−0.217	0.081	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__21DPE	0.045	−0.080	0.171	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.011	−0.156	0.134	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.069	−0.214	0.076	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.023	−0.172	0.125	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.034	−0.179	0.110	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__21DPE	−0.008	−0.133	0.118	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.047	−0.196	0.102	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.056	−0.205	0.093	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.019	−0.168	0.130	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.095	−0.059	0.248	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	−0.024	−0.173	0.125	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__28DPE	0.089	−0.036	0.215	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.033	−0.111	0.178	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	−0.025	−0.170	0.120	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	0.021	−0.128	0.169	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	0.010	−0.135	0.154	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__28DPE	0.036	−0.089	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.003	−0.152	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.012	−0.161	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	0.025	−0.124	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.139	−0.015	0.292	0.125	ns
Distance	Temp.DPE	32°C__0DPE	28°C__42DPE	0.114	−0.016	0.243	0.161	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.057	−0.091	0.206	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.001	−0.150	0.148	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	0.045	−0.108	0.197	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	0.034	−0.115	0.183	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__42DPE	0.061	−0.069	0.191	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	0.021	−0.131	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	0.012	−0.140	0.165	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	0.049	−0.103	0.202	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	0.163	0.006	0.320	0.034	*
Distance	Temp.DPE	32°C__14DPE	32°C__0DPE	−0.056	−0.182	0.069	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__0DPE	−0.114	−0.240	0.011	0.115	ns
Distance	Temp.DPE	32°C__28DPE	32°C__0DPE	−0.069	−0.199	0.061	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__0DPE	−0.080	−0.205	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__0DPE	−0.053	−0.155	0.049	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__0DPE	−0.092	−0.222	0.037	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__0DPE	−0.101	−0.231	0.029	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__0DPE	−0.064	−0.194	0.066	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__0DPE	0.049	−0.086	0.185	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.058	−0.203	0.086	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.013	−0.161	0.136	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.024	−0.168	0.121	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__14DPE	0.003	−0.122	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.036	−0.185	0.112	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.045	−0.194	0.104	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.008	−0.157	0.141	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	0.105	−0.048	0.259	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	0.046	−0.103	0.194	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	0.035	−0.110	0.179	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__21DPE	0.061	−0.064	0.187	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	0.022	−0.127	0.171	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	0.013	−0.136	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	0.050	−0.099	0.199	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	0.164	0.010	0.317	0.025	*
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.011	−0.160	0.138	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__28DPE	0.016	−0.114	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.024	−0.176	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.033	−0.185	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	0.004	−0.148	0.157	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	0.118	−0.039	0.275	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__42DPE	0.027	−0.099	0.152	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.013	−0.161	0.136	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.021	−0.170	0.127	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	0.015	−0.133	0.164	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	0.129	−0.024	0.283	0.209	ns
Distance	Temp.DPE	35°C__14DPE	35°C__0DPE	−0.039	−0.169	0.090	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__0DPE	−0.048	−0.178	0.082	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__0DPE	−0.011	−0.141	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__0DPE	0.102	−0.033	0.238	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.009	−0.161	0.144	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	0.028	−0.124	0.181	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.142	−0.016	0.299	0.127	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	0.037	−0.116	0.190	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.151	−0.007	0.308	0.076	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.114	−0.044	0.271	≥0.25	ns
canberra
Distance	Temp.DPE	28°C__14DPE	28°C__0DPE	−0.033	−0.138	0.072	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__0DPE	−0.005	−0.103	0.092	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__0DPE	−0.074	−0.171	0.024	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__0DPE	−0.069	−0.170	0.032	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__0DPE	0.035	−0.045	0.115	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__0DPE	−0.021	−0.119	0.076	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__0DPE	−0.070	−0.167	0.028	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__0DPE	−0.056	−0.157	0.045	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__0DPE	−0.068	−0.165	0.030	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__0DPE	0.008	−0.071	0.088	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__0DPE	−0.041	−0.142	0.060	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__0DPE	−0.047	−0.148	0.054	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__0DPE	−0.052	−0.153	0.049	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__0DPE	0.045	−0.061	0.150	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	0.028	−0.091	0.147	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.040	−0.160	0.079	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.035	−0.158	0.087	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__14DPE	0.068	−0.037	0.174	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	0.012	−0.107	0.131	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.037	−0.156	0.083	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.023	−0.145	0.099	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.034	−0.154	0.085	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__14DPE	0.042	−0.064	0.147	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.008	−0.130	0.115	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.013	−0.136	0.109	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.019	−0.141	0.103	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	0.078	−0.048	0.204	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.068	−0.181	0.044	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	−0.063	−0.179	0.052	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__21DPE	0.040	−0.057	0.138	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.016	−0.128	0.097	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.065	−0.177	0.048	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.051	−0.167	0.065	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.062	−0.175	0.050	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__21DPE	0.014	−0.084	0.111	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.036	−0.151	0.080	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.041	−0.157	0.074	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.047	−0.162	0.069	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.050	−0.070	0.169	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.005	−0.110	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__28DPE	0.109	0.011	0.206	0.014	*
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.052	−0.060	0.165	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	0.004	−0.109	0.116	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	0.017	−0.098	0.133	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	0.006	−0.106	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__28DPE	0.082	−0.015	0.180	0.206	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	0.033	−0.083	0.148	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	0.027	−0.088	0.143	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	0.021	−0.094	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.118	−0.001	0.237	0.055	ns
Distance	Temp.DPE	32°C__0DPE	28°C__42DPE	0.104	0.003	0.205	0.038	*
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.047	−0.068	0.163	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.001	−0.117	0.114	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	0.012	−0.106	0.131	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	0.001	−0.115	0.117	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__42DPE	0.077	−0.024	0.178	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	0.028	−0.091	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	0.022	−0.097	0.141	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	0.016	−0.102	0.135	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	0.113	−0.009	0.235	0.104	ns
Distance	Temp.DPE	32°C__14DPE	32°C__0DPE	−0.056	−0.154	0.041	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__0DPE	−0.105	−0.202	−0.007	0.022	*
Distance	Temp.DPE	32°C__28DPE	32°C__0DPE	−0.091	−0.192	0.010	0.125	ns
Distance	Temp.DPE	32°C__42DPE	32°C__0DPE	−0.103	−0.200	−0.005	0.029	*
Distance	Temp.DPE	35°C__0DPE	32°C__0DPE	−0.027	−0.106	0.053	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__0DPE	−0.076	−0.177	0.025	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__0DPE	−0.082	−0.183	0.019	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__0DPE	−0.087	−0.188	0.014	0.175	ns
Distance	Temp.DPE	35°C__42DPE	32°C__0DPE	0.009	−0.096	0.115	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.049	−0.161	0.064	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.035	−0.151	0.081	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.046	−0.159	0.066	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__14DPE	0.030	−0.068	0.127	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.020	−0.135	0.096	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.025	−0.141	0.090	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.031	−0.147	0.085	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	0.066	−0.054	0.185	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	0.014	−0.102	0.129	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	0.002	−0.110	0.115	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__21DPE	0.078	−0.019	0.176	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	0.029	−0.087	0.145	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	0.023	−0.092	0.139	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	0.018	−0.098	0.133	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	0.114	−0.005	0.234	0.076	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.011	−0.127	0.104	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__28DPE	0.065	−0.036	0.166	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	0.015	−0.103	0.134	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	0.010	−0.109	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	0.004	−0.115	0.123	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	0.101	−0.022	0.223	0.237	ns
Distance	Temp.DPE	35°C__0DPE	32°C__42DPE	0.076	−0.021	0.173	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	0.027	−0.089	0.142	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	0.021	−0.095	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	0.015	−0.100	0.131	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	0.112	−0.007	0.231	0.091	ns
Distance	Temp.DPE	35°C__14DPE	35°C__0DPE	−0.049	−0.150	0.052	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__0DPE	−0.055	−0.156	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__0DPE	−0.061	−0.162	0.040	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__0DPE	0.036	−0.069	0.141	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.006	−0.124	0.113	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.011	−0.130	0.107	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.085	−0.037	0.208	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.006	−0.124	0.113	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.091	−0.031	0.213	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.097	−0.026	0.219	≥0.25	ns
gunifrac
Distance	Temp.DPE	28°C__14DPE	28°C__0DPE	−0.005	−0.116	0.107	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__0DPE	0.038	−0.065	0.141	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__0DPE	−0.010	−0.113	0.093	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__0DPE	−0.033	−0.140	0.073	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__0DPE	0.067	−0.017	0.151	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__0DPE	0.018	−0.085	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__0DPE	−0.023	−0.126	0.080	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__0DPE	−0.014	−0.121	0.093	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__0DPE	−0.026	−0.129	0.077	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__0DPE	0.029	−0.055	0.113	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__0DPE	−0.015	−0.122	0.092	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__0DPE	−0.020	−0.127	0.087	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__0DPE	−0.065	−0.171	0.042	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__0DPE	0.091	−0.020	0.202	0.245	ns
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	0.042	−0.084	0.168	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.005	−0.131	0.121	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.029	−0.158	0.100	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__14DPE	0.072	−0.040	0.183	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	0.022	−0.104	0.148	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.019	−0.145	0.107	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.009	−0.138	0.120	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.021	−0.147	0.105	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__14DPE	0.033	−0.078	0.145	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.010	−0.139	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.015	−0.145	0.114	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.060	−0.189	0.069	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	0.096	−0.037	0.229	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.047	−0.166	0.072	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	−0.071	−0.193	0.051	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__21DPE	0.029	−0.074	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.020	−0.139	0.099	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.061	−0.180	0.058	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.051	−0.174	0.071	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.063	−0.182	0.056	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__21DPE	−0.009	−0.112	0.094	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.052	−0.175	0.070	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.058	−0.180	0.065	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.102	−0.224	0.020	0.216	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.054	−0.073	0.180	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	−0.024	−0.146	0.098	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__28DPE	0.077	−0.026	0.180	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.027	−0.092	0.146	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	−0.014	−0.133	0.105	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	−0.004	−0.126	0.118	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	−0.016	−0.135	0.103	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__28DPE	0.038	−0.065	0.142	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.005	−0.127	0.117	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.010	−0.132	0.112	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.055	−0.177	0.067	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.101	−0.025	0.227	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__42DPE	0.100	−0.006	0.207	0.089	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.051	−0.071	0.173	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	0.010	−0.112	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	0.020	−0.106	0.145	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	0.008	−0.114	0.130	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__42DPE	0.062	−0.045	0.169	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	0.019	−0.107	0.144	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	0.014	−0.112	0.139	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.031	−0.157	0.094	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	0.125	−0.005	0.254	0.071	ns
Distance	Temp.DPE	32°C__14DPE	32°C__0DPE	−0.049	−0.152	0.054	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__0DPE	−0.090	−0.193	0.013	0.155	ns
Distance	Temp.DPE	32°C__28DPE	32°C__0DPE	−0.081	−0.188	0.026	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__0DPE	−0.093	−0.196	0.010	0.130	ns
Distance	Temp.DPE	35°C__0DPE	32°C__0DPE	−0.038	−0.122	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__0DPE	−0.082	−0.189	0.025	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__0DPE	−0.087	−0.194	0.020	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__0DPE	−0.132	−0.238	−0.025	0.003	**
Distance	Temp.DPE	35°C__42DPE	32°C__0DPE	0.024	−0.087	0.135	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.041	−0.160	0.078	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.031	−0.154	0.091	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.043	−0.162	0.076	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__14DPE	0.011	−0.092	0.114	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.032	−0.155	0.090	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.037	−0.160	0.085	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.082	−0.204	0.040	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	0.074	−0.053	0.200	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	0.010	−0.113	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.002	−0.121	0.117	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__21DPE	0.052	−0.051	0.155	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	0.009	−0.114	0.131	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	0.004	−0.119	0.126	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.041	−0.163	0.081	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	0.115	−0.011	0.241	0.119	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.012	−0.134	0.110	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__28DPE	0.043	−0.064	0.149	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.001	−0.126	0.124	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.006	−0.132	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.051	−0.176	0.075	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	0.105	−0.024	0.234	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__42DPE	0.054	−0.049	0.157	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	0.011	−0.111	0.133	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	0.006	−0.117	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.039	−0.161	0.083	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	0.117	−0.009	0.243	0.102	ns
Distance	Temp.DPE	35°C__14DPE	35°C__0DPE	−0.044	−0.150	0.063	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__0DPE	−0.049	−0.155	0.058	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__0DPE	−0.093	−0.200	0.013	0.160	ns
Distance	Temp.DPE	35°C__42DPE	35°C__0DPE	0.062	−0.049	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.005	−0.130	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.050	−0.175	0.076	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.106	−0.023	0.235	0.244	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.045	−0.170	0.081	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.111	−0.018	0.240	0.180	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.156	0.027	0.285	0.005	**

Addl. Metrics

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S2A

S2B

S2C

S2D

Fig. 3) Infection burden is highest in fish reared at lower water temperatures

Fig. 3 Infection outcomes in zebrafish exposed to Pseudocapillaria tomentosa. (A) Histological sections stained with ### stain in zebrafish exposed to P. tomentosa examined at ### days post exposure. Nematodes in intestinal lumen. Arrow = ###. (B) Infection outcome analysis of fish exposed to P. tomentosa (n = 89) by temperature. Fish reared at 28°C and 32°C water temperatures had significantly different infection burden to fish reared at 35°C water temperature. Only one fish reared at 35°C was identified as being positively infected by wet mount. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level.

3A)

3B)

Tables

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GLM

GLM Results
glm.nb(Total.Worm.Count ~ Temperature); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
(Intercept)	1.552	0.316	4.914	<0.001	****
Temperature32	−0.368	0.431	−0.853	≥0.25	ns
Temperature35	−5.078	1.080	−4.701	<0.001	****

ANOVA

ANOVA of GLM
ANOVA(GLM.NB(Total.Worm.Count ~ Temperature), type = 2); Exposed fish
term	statistic	df	p.value
Temperature	55.264	2.000	<0.001

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Temperature); Exposed fish
term	.y.	group1	group2	estimate	std.error	df	statistic	adj.p.value
Temperature	Total.Worm.Count	28	32	0.368	0.431	Inf	0.853	≥0.25
Temperature	Total.Worm.Count	28	35	5.078	1.080	Inf	4.701	<0.001
Temperature	Total.Worm.Count	32	35	4.710	1.074	Inf	4.386	<0.001

(hide)

Pathology Results

Summary of Infection Outcomes
(Pathology Results)
Pathology.Results	Count
28
negative	8
positive	17
32
negative	15
positive	15
35
negative	33
positive	1

Total Worm Counts

Summary of Infection Outcomes
(Total Worm Count)
Group	Variable	Mean	SD	Min	Max	n_Missing
28	Total.Worm.Count	5	5	0	14	0
32	Total.Worm.Count	3	5	0	15	0
35	Total.Worm.Count	0	0	0	1	0

Suppl.

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S3B.1

Tables

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GLM

GLM Results
glm.nb(Total.Worm.Count ~ Temperature*DPE); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
(Intercept)	0.535	0.795	0.673	≥0.25	ns
Temperature32	0.233	1.081	0.216	≥0.25	ns
Temperature35	−4.474	2.964	−1.509	0.131	ns
DPE	0.035	0.027	1.304	0.192	ns
Temperature32:DPE	−0.019	0.038	−0.501	≥0.25	ns
Temperature35:DPE	−0.020	0.099	−0.200	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM.NB(Total.Worm.Count ~ Temperature*DPE), type = 2); Exposed fish
term	statistic	df	p.value
Temperature	56.284	2.000	<0.001
DPE	1.790	1.000	0.181
Temperature:DPE	0.265	2.000	≥0.25

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Temperature*DPE); Exposed fish
DPE	.y.	term	group1	group2	estimate	std.error	df	statistic	adj.p.value
14	Total.Worm.Count	Temperature	28	32	−0.229	0.279	Inf	−0.820	≥0.25
14	Total.Worm.Count	Temperature	28	35	38.009	22369621.333	Inf	0.000	≥0.25
14	Total.Worm.Count	Temperature	32	35	38.238	22369621.333	Inf	0.000	≥0.25
21	Total.Worm.Count	Temperature	28	32	0.560	0.392	Inf	1.427	≥0.25
21	Total.Worm.Count	Temperature	28	35	38.222	23726566.406	Inf	0.000	≥0.25
21	Total.Worm.Count	Temperature	32	35	37.663	23726566.406	Inf	0.000	≥0.25
28	Total.Worm.Count	Temperature	28	32	0.605	0.582	Inf	1.040	≥0.25
28	Total.Worm.Count	Temperature	28	35	3.837	1.121	Inf	3.422	0.002
28	Total.Worm.Count	Temperature	32	35	3.232	1.119	Inf	2.888	0.011
42	Total.Worm.Count	Temperature	28	32	0.377	0.620	Inf	0.609	≥0.25
42	Total.Worm.Count	Temperature	28	35	21.230	3142.206	Inf	0.007	≥0.25
42	Total.Worm.Count	Temperature	32	35	20.853	3142.206	Inf	0.007	≥0.25

S3B.2

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GLM

GLM Results
glm.nb(Total.Worm.Count ~ Temperature*DPE); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
(Intercept)	0.535	0.795	0.673	≥0.25	ns
Temperature32	0.233	1.081	0.216	≥0.25	ns
Temperature35	−4.474	2.964	−1.509	0.131	ns
DPE	0.035	0.027	1.304	0.192	ns
Temperature32:DPE	−0.019	0.038	−0.501	≥0.25	ns
Temperature35:DPE	−0.020	0.099	−0.200	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM.NB(Total.Worm.Count ~ Temperature*DPE), type = 2); Exposed fish
term	statistic	df	p.value
Temperature	56.284	2.000	<0.001
DPE	1.790	1.000	0.181
Temperature:DPE	0.265	2.000	≥0.25

S3C

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Counts

Detection Method Comparison by DPE
Number of positive and negative detections for Wet and Histo methods
Days Post Exposure	Histo Positive	Wet Positive	Histo Negative	Wet Negative
28
14	9	6	0	4
21	6	6	2	4
28	6	8	0	0
42	8	7	1	2
32
14	6	4	3	6
21	5	5	0	5
28	7	7	1	2
42	7	7	2	2
35
14	1	1	9	9
21	6	0	3	10
28	1	1	2	7
42	1	0	9	10
NA
NA	0	0	0	0

Stats

Statistical Comparison of Detection Methods
McNemar's test comparing Wet and Histo methods by Temperature and DPE
Days Post Exposure	Histo Only	Wet Only	Both Positive	Both Negative	P-value1	Chi-squared
28
14	3	0	6	0	0.2482	1.333333
21	1	0	5	2	1.0000	0.000000
28	0	0	6	0	NaN	NaN
42	1	0	6	1	1.0000	0.000000
32
14	4	1	2	2	0.3711	0.800000
21	2	0	3	0	0.4795	0.500000
28	1	0	6	1	1.0000	0.000000
42	2	2	5	0	1.0000	0.000000
35
14	1	1	0	8	1.0000	0.000000
21	6	0	0	3	0.0412	4.166667
28	0	0	1	2	NaN	NaN
42	1	0	0	9	1.0000	0.000000
NA
NA	0	0	0	0	NaN	NaN
1 McNemar's test compares the discordant pairs (Wet Only vs Histo Only)

Fig. 4) Gut microbiome response to parasite exposure varies across water temperature

Fig. 4 Effects of Pseudocapillaria tomentosa exposure on zebrafish gut microbiomes reared at different water temperatures. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C water temperature to fish reared at 32°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main effect of temperature. The analysis shows that gut microbiome composition significantly differs between parasite exposed fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity decreases with time from 0 days post exposure (dpe) to 42 dpe in parasite exposed fish reared at 28°C water temperature. (D) Capscale ordination of gut microbiome composition based on the Canberra dissimilarity constrained on the main effects of water temperature and time (days post exposure, dpe), and their interaction. The analysis shows that shows that gut microbiome composition differs between parasite exposed fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated covariates.

4A)

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GLM

GLM Results
glm(Alpha.Score ~ Temperature); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.936	0.233	−4.021	<0.001	***
Temperature32	0.909	0.301	3.017	0.003	**
Temperature35	0.516	0.296	1.741	0.085	ns
Simpson
(Intercept)	−1.002	0.243	−4.130	<0.001	****
Temperature32	1.057	0.312	3.384	0.001	**
Temperature35	0.772	0.306	2.525	0.013	*
Richness
(Intercept)	−0.681	0.190	−3.583	<0.001	***
Temperature32	0.703	0.251	2.800	0.006	**
Temperature35	0.242	0.247	0.978	≥0.25	ns
Phylogenetic
(Intercept)	−0.681	0.180	−3.789	<0.001	***
Temperature32	0.648	0.237	2.730	0.008	**
Temperature35	0.161	0.235	0.688	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	9.440	2.000	0.009	**
Simpson
Temperature	12.497	2.000	0.002	**
Richness
Temperature	8.575	2.000	0.014	*
Phylogenetic
Temperature	8.706	2.000	0.013	*

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Exposed fish
term	.y.	group1	group2	estimate	std.error	statistic	adj.p.value	Variable
Shannon
Temperature	Alpha.Score	32	28	0.909	0.301	3.017	0.007	Temperature
Temperature	Alpha.Score	35	28	0.516	0.296	1.741	0.190	Temperature
Temperature	Alpha.Score	35	32	−0.393	0.265	−1.482	≥0.25	Temperature
Simpson
Temperature	Alpha.Score	32	28	1.057	0.312	3.384	0.002	Temperature
Temperature	Alpha.Score	35	28	0.772	0.306	2.525	0.031	Temperature
Temperature	Alpha.Score	35	32	−0.285	0.270	−1.054	≥0.25	Temperature
Richness
Temperature	Alpha.Score	32	28	0.703	0.251	2.800	0.014	Temperature
Temperature	Alpha.Score	35	28	0.242	0.247	0.978	≥0.25	Temperature
Temperature	Alpha.Score	35	32	−0.461	0.228	−2.028	0.105	Temperature
Phylogenetic
Temperature	Alpha.Score	32	28	0.648	0.237	2.730	0.017	Temperature
Temperature	Alpha.Score	35	28	0.161	0.235	0.688	≥0.25	Temperature
Temperature	Alpha.Score	35	32	−0.487	0.216	−2.251	0.063	Temperature

Addl. Metrics

4B)

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.447	0.001	***
Residual	86.000	15.530	0.749	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.701	0.001	***
Residual	86.000	26.947	0.883	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.590	0.001	***
Residual	86.000	12.478	0.788	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temperature	2.000	0.364	0.182	8.708	<0.001	***
Residual	86.000	1.796	0.021	NA	NA	NA
canberra
Temperature	2.000	0.314	0.157	17.419	<0.001	****
Residual	86.000	0.774	0.009	NA	NA	NA
gunifrac
Temperature	2.000	0.310	0.155	13.567	<0.001	****
Residual	86.000	0.983	0.011	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temperature	32	28	−0.043	−0.136	0.051	≥0.25	ns
Distance	Temperature	35	28	−0.150	−0.241	−0.059	<0.001	***
Distance	Temperature	35	32	−0.108	−0.194	−0.021	0.011	*
canberra
Distance	Temperature	32	28	−0.110	−0.171	−0.049	<0.001	***
Distance	Temperature	35	28	−0.144	−0.204	−0.085	<0.001	****
Distance	Temperature	35	32	−0.034	−0.091	0.022	≥0.25	ns
gunifrac
Distance	Temperature	32	28	−0.056	−0.125	0.013	0.133	ns
Distance	Temperature	35	28	−0.143	−0.211	−0.076	<0.001	****
Distance	Temperature	35	32	−0.087	−0.151	−0.023	0.005	**

Addl. Metrics

4C)

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*DPE); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.134	0.594	−0.226	≥0.25	ns
Temperature32	0.228	0.774	0.295	≥0.25	ns
Temperature35	−0.428	0.777	−0.551	≥0.25	ns
DPE	−0.031	0.022	−1.425	0.158	ns
Temperature32:DPE	0.026	0.028	0.917	≥0.25	ns
Temperature35:DPE	0.036	0.028	1.300	0.197	ns
Simpson
(Intercept)	−0.282	0.617	−0.456	≥0.25	ns
Temperature32	0.336	0.801	0.420	≥0.25	ns
Temperature35	−0.172	0.799	−0.215	≥0.25	ns
DPE	−0.028	0.022	−1.234	0.221	ns
Temperature32:DPE	0.028	0.029	0.946	≥0.25	ns
Temperature35:DPE	0.036	0.029	1.261	0.211	ns
Richness
(Intercept)	0.316	0.480	0.657	≥0.25	ns
Temperature32	−0.034	0.636	−0.054	≥0.25	ns
Temperature35	−0.596	0.638	−0.934	≥0.25	ns
DPE	−0.038	0.017	−2.191	0.031	*
Temperature32:DPE	0.028	0.023	1.199	0.234	ns
Temperature35:DPE	0.032	0.023	1.405	0.164	ns
Phylogenetic
(Intercept)	0.305	0.451	0.676	≥0.25	ns
Temperature32	−0.101	0.597	−0.169	≥0.25	ns
Temperature35	−0.766	0.601	−1.274	0.206	ns
DPE	−0.038	0.016	−2.310	0.023	*
Temperature32:DPE	0.028	0.022	1.298	0.198	ns
Temperature35:DPE	0.036	0.022	1.651	0.103	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	8.789	2.000	0.012	*
DPE	0.523	1.000	≥0.25	ns
Temperature:DPE	1.759	2.000	≥0.25	ns
Simpson
Temperature	11.871	2.000	0.003	**
DPE	0.120	1.000	≥0.25	ns
Temperature:DPE	1.684	2.000	≥0.25	ns
Richness
Temperature	7.911	2.000	0.019	*
DPE	3.435	1.000	0.064	ns
Temperature:DPE	2.255	2.000	≥0.25	ns
Phylogenetic
Temperature	8.173	2.000	0.017	*
DPE	3.103	1.000	0.078	ns
Temperature:DPE	2.980	2.000	0.225	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	DPE	21	14	−3.311	1.822	−1.817	0.246	DPE	28
28	Alpha.Score	DPE	28	14	−2.027	1.023	−1.982	0.178	DPE	28
28	Alpha.Score	DPE	42	14	−1.024	0.705	−1.452	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	1.284	1.982	0.648	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	2.287	1.838	1.245	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	1.004	1.050	0.956	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.536	0.537	−0.999	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.056	0.558	0.099	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.271	0.532	−0.509	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.592	0.606	0.977	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.266	0.581	0.457	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	−0.326	0.601	−0.543	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.246	0.270	−0.909	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.292	0.271	−1.075	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.097	0.258	0.377	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	−0.046	0.283	−0.162	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.343	0.270	1.273	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.389	0.271	1.438	≥0.25	DPE	35
Simpson
28	Alpha.Score	DPE	21	14	−3.141	1.820	−1.725	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	−1.994	1.072	−1.861	0.227	DPE	28
28	Alpha.Score	DPE	42	14	−0.925	0.721	−1.283	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	1.147	2.001	0.573	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	2.215	1.837	1.206	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	1.069	1.100	0.971	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.416	0.550	−0.756	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.275	0.580	0.474	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.138	0.546	−0.252	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.691	0.626	1.103	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.278	0.595	0.467	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	−0.413	0.623	−0.662	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.374	0.297	−1.258	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.285	0.296	−0.966	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.142	0.284	0.502	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	0.088	0.308	0.287	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.516	0.297	1.738	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.428	0.295	1.448	≥0.25	DPE	35
Richness
28	Alpha.Score	DPE	21	14	−2.219	1.016	−2.185	0.123	DPE	28
28	Alpha.Score	DPE	28	14	−1.585	0.795	−1.993	0.184	DPE	28
28	Alpha.Score	DPE	42	14	−1.239	0.650	−1.907	0.218	DPE	28
28	Alpha.Score	DPE	28	21	0.634	1.133	0.560	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	0.980	1.036	0.946	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	0.346	0.821	0.421	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.561	0.412	−1.363	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	−0.470	0.430	−1.091	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.357	0.410	−0.871	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.092	0.462	0.199	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.204	0.444	0.460	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	0.112	0.461	0.243	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.338	0.242	−1.396	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.329	0.242	−1.361	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	−0.233	0.233	−1.000	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	0.009	0.252	0.035	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.105	0.244	0.430	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.096	0.244	0.394	≥0.25	DPE	35
Phylogenetic
28	Alpha.Score	DPE	21	14	−2.030	0.921	−2.205	0.118	DPE	28
28	Alpha.Score	DPE	28	14	−1.472	0.742	−1.985	0.188	DPE	28
28	Alpha.Score	DPE	42	14	−1.216	0.616	−1.975	0.192	DPE	28
28	Alpha.Score	DPE	28	21	0.557	1.029	0.542	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	0.814	0.942	0.864	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	0.256	0.768	0.334	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.537	0.382	−1.406	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	−0.442	0.399	−1.108	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.329	0.380	−0.866	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.095	0.430	0.222	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.208	0.413	0.504	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	0.113	0.428	0.263	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.273	0.236	−1.159	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.214	0.235	−0.911	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	−0.121	0.226	−0.535	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	0.059	0.246	0.242	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.152	0.237	0.642	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.093	0.236	0.393	≥0.25	DPE	35

Addl. Metrics

4D)

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.843	0.001	***
DPE	1.000	0.394	0.019	2.241	0.056	ns
Temperature:DPE	2.000	0.548	0.026	1.558	0.113	ns
Residual	83.000	14.589	0.703	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.847	0.001	***
DPE	1.000	0.674	0.022	2.207	0.006	**
Temperature:DPE	2.000	0.913	0.030	1.494	0.017	*
Residual	83.000	25.360	0.831	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.847	0.001	***
DPE	1.000	0.261	0.016	1.837	0.101	ns
Temperature:DPE	2.000	0.436	0.028	1.535	0.102	ns
Residual	83.000	11.781	0.744	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.DPE	11.000	0.624	0.057	3.261	0.001	**
Residual	77.000	1.339	0.017	NA	NA	NA
canberra
Temp.DPE	11.000	0.316	0.029	2.748	0.005	**
Residual	77.000	0.805	0.010	NA	NA	NA
gunifrac
Temp.DPE	11.000	0.420	0.038	3.232	0.001	**
Residual	77.000	0.909	0.012	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	−0.213	−0.485	0.059	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.258	−0.512	−0.005	0.042	*
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.063	−0.285	0.160	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	−0.059	−0.270	0.152	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.089	−0.319	0.141	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.123	−0.363	0.117	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.142	−0.372	0.088	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.202	−0.418	0.014	0.090	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.170	−0.393	0.052	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.289	−0.511	−0.067	0.002	**
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	−0.192	−0.408	0.024	0.130	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.046	−0.344	0.253	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	0.150	−0.122	0.422	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	0.153	−0.110	0.416	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	0.124	−0.154	0.403	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	0.090	−0.197	0.377	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	0.071	−0.208	0.350	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	0.011	−0.256	0.278	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	0.042	−0.230	0.315	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.077	−0.349	0.196	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.021	−0.246	0.288	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.196	−0.058	0.449	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.199	−0.044	0.443	0.220	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	0.170	−0.091	0.430	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	0.136	−0.133	0.405	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	0.117	−0.144	0.377	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	0.057	−0.191	0.305	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	0.088	−0.165	0.341	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.031	−0.284	0.222	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.066	−0.182	0.314	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.003	−0.207	0.214	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.026	−0.256	0.204	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	−0.060	−0.300	0.180	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	−0.079	−0.309	0.151	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	−0.139	−0.355	0.077	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	−0.108	−0.330	0.115	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.227	−0.449	−0.004	0.042	*
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	−0.129	−0.345	0.087	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.029	−0.248	0.190	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.063	−0.293	0.166	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.082	−0.301	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.142	−0.347	0.062	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.111	−0.322	0.100	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.230	−0.441	−0.019	0.021	*
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	−0.133	−0.337	0.072	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	−0.034	−0.281	0.213	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.053	−0.291	0.185	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	−0.113	−0.337	0.111	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	−0.082	−0.312	0.148	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.201	−0.431	0.029	0.148	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	−0.103	−0.327	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.019	−0.266	0.228	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.079	−0.313	0.155	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.048	−0.288	0.192	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.167	−0.407	0.073	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	−0.069	−0.304	0.165	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.060	−0.284	0.164	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.029	−0.259	0.201	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.148	−0.378	0.082	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	−0.050	−0.274	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	0.031	−0.185	0.247	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.088	−0.304	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.010	−0.200	0.219	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.119	−0.341	0.103	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	−0.022	−0.238	0.194	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.097	−0.119	0.313	≥0.25	ns
canberra
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	−0.004	−0.215	0.207	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	0.040	−0.156	0.237	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	0.059	−0.113	0.232	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	−0.043	−0.206	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.044	−0.223	0.134	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.102	−0.288	0.084	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.084	−0.262	0.095	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.097	−0.265	0.070	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.097	−0.269	0.075	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.142	−0.314	0.031	0.215	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	−0.103	−0.271	0.064	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	0.044	−0.187	0.276	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	0.064	−0.148	0.275	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.039	−0.243	0.165	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.040	−0.256	0.176	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.097	−0.320	0.125	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.079	−0.295	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.093	−0.300	0.114	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.093	−0.304	0.118	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.137	−0.348	0.074	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	−0.099	−0.306	0.108	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.019	−0.177	0.216	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	−0.083	−0.272	0.106	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	−0.085	−0.286	0.117	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	−0.142	−0.351	0.067	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	−0.124	−0.326	0.078	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.137	−0.329	0.055	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.137	−0.334	0.059	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.182	−0.378	0.015	0.097	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	−0.144	−0.336	0.049	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	−0.102	−0.266	0.061	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.104	−0.282	0.075	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	−0.161	−0.347	0.025	0.157	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	−0.143	−0.321	0.036	0.247	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	−0.156	−0.324	0.011	0.091	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	−0.156	−0.329	0.016	0.112	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.201	−0.373	−0.028	0.010	**
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	−0.163	−0.330	0.005	0.065	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.001	−0.171	0.168	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.059	−0.237	0.119	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.041	−0.211	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.054	−0.212	0.104	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.054	−0.218	0.109	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.099	−0.262	0.065	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	−0.060	−0.219	0.098	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	−0.057	−0.249	0.134	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.039	−0.224	0.145	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	−0.053	−0.226	0.121	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	−0.053	−0.231	0.126	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.097	−0.276	0.081	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	−0.059	−0.233	0.115	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	0.018	−0.174	0.210	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	0.005	−0.177	0.186	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	0.005	−0.182	0.191	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.040	−0.226	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	−0.002	−0.183	0.180	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.013	−0.187	0.160	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.014	−0.192	0.165	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.058	−0.236	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	−0.020	−0.194	0.154	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	0.000	−0.168	0.167	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.044	−0.212	0.123	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	−0.006	−0.169	0.156	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.044	−0.217	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	−0.006	−0.174	0.161	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.038	−0.129	0.206	≥0.25	ns
gunifrac
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	−0.191	−0.415	0.033	0.173	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.061	−0.270	0.147	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	0.018	−0.165	0.201	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	−0.024	−0.197	0.150	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.027	−0.216	0.163	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.100	−0.298	0.098	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.069	−0.259	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.131	−0.309	0.047	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.137	−0.320	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.198	−0.382	−0.015	0.022	*
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	−0.134	−0.312	0.044	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	0.130	−0.116	0.375	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	0.209	−0.015	0.433	0.092	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	0.167	−0.049	0.384	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	0.164	−0.065	0.394	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	0.091	−0.146	0.327	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	0.122	−0.108	0.351	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	0.060	−0.161	0.280	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	0.054	−0.170	0.278	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.007	−0.232	0.217	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.057	−0.163	0.277	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.079	−0.130	0.288	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.038	−0.163	0.238	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	0.035	−0.180	0.249	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	−0.039	−0.260	0.183	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	−0.008	−0.222	0.206	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.070	−0.274	0.134	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.075	−0.284	0.133	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.137	−0.346	0.072	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	−0.072	−0.277	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	−0.041	−0.215	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.045	−0.234	0.145	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	−0.118	−0.316	0.080	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	−0.087	−0.277	0.102	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	−0.149	−0.327	0.029	0.190	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	−0.155	−0.338	0.029	0.182	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.216	−0.399	−0.033	0.008	**
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	−0.151	−0.329	0.026	0.173	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.003	−0.184	0.177	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.076	−0.266	0.113	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.046	−0.226	0.135	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.108	−0.276	0.060	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.113	−0.287	0.061	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.175	−0.348	−0.001	0.047	*
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	−0.110	−0.278	0.058	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	−0.073	−0.277	0.130	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.043	−0.238	0.153	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	−0.105	−0.289	0.080	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	−0.110	−0.300	0.079	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.172	−0.361	0.018	0.113	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	−0.107	−0.292	0.078	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	0.031	−0.173	0.234	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.031	−0.224	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.037	−0.234	0.161	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.098	−0.296	0.099	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	−0.034	−0.227	0.159	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.062	−0.247	0.122	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.067	−0.257	0.122	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.129	−0.319	0.060	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	−0.064	−0.249	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.005	−0.183	0.173	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.067	−0.245	0.111	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	−0.002	−0.175	0.170	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.062	−0.245	0.121	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.003	−0.175	0.181	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.065	−0.113	0.243	≥0.25	ns

Addl. Metrics

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S4A

S4B

S4C

S4D

Fig. 5) Gut microbiome response has a non-linear relationship with infection burden

Fig. 5 The impacts of presence of infection and infection burden on the gut microbiomes of Pseudocapillaria tomentosa exposed zebrafish. (A) Simpson’s Index for diversity of parasite exposed fish. Gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature depending on presence of infection. (B) Capscale ordination based on the Canberra dissimilarity of gut microbiome composition of parasite exposed fish constrained on the main effects of temperature and pathology result. The analysis shows that gut microbiome composition significantly differs between positively infected fish reared at different water temperatures. (C) Infection burden (total worm counts ) is positively correlated with lowest or highest alpha diversity scores in positively infected fish. (D) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main effects of water temperature and infection burden. The analysis shows that gut microbiome composition significantly differs between clusters of Low, High and Other fish. Samples points are colored by water temperature, and filled by cluster grouping. Samples with at least one detectable worm and an alpha-diversity score less than 0.5 are categorized as Low (orange fill), samples with at least one detectable worm and an alpha-diversity score greater than 0.5 are categorized as High (purple fill), and samples with no observable infection are categorized as Other (white and transparent fill). Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

5A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*Pathology.Results); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−1.090	0.429	−2.544	0.013	*
Temperature32	0.889	0.508	1.749	0.084	ns
Temperature35	0.693	0.468	1.482	0.142	ns
Pathology.Resultspositive	0.223	0.512	0.436	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.125	0.641	0.195	≥0.25	ns
Temperature35:Pathology.Resultspositive	−1.122	1.392	−0.806	≥0.25	ns
Simpson
(Intercept)	−1.265	0.458	−2.760	0.007	**
Temperature32	1.105	0.536	2.061	0.042	*
Temperature35	1.055	0.495	2.130	0.036	*
Pathology.Resultspositive	0.375	0.541	0.694	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.054	0.669	0.081	≥0.25	ns
Temperature35:Pathology.Resultspositive	−1.154	1.337	−0.863	≥0.25	ns
Richness
(Intercept)	−0.421	0.329	−1.279	0.204	ns
Temperature32	0.483	0.405	1.194	0.236	ns
Temperature35	−0.002	0.367	−0.005	≥0.25	ns
Pathology.Resultspositive	−0.390	0.407	−0.957	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.310	0.526	0.589	≥0.25	ns
Temperature35:Pathology.Resultspositive	−0.221	1.125	−0.197	≥0.25	ns
Phylogenetic
(Intercept)	−0.421	0.311	−1.353	0.180	ns
Temperature32	0.397	0.383	1.037	≥0.25	ns
Temperature35	−0.085	0.348	−0.244	≥0.25	ns
Pathology.Resultspositive	−0.390	0.385	−1.014	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.373	0.497	0.750	≥0.25	ns
Temperature35:Pathology.Resultspositive	−0.112	1.058	−0.105	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	9.895	2.000	0.007	**
Pathology.Results	0.569	1.000	≥0.25	ns
Temperature:Pathology.Results	0.997	2.000	≥0.25	ns
Simpson
Temperature	13.663	2.000	0.001	**
Pathology.Results	1.134	1.000	≥0.25	ns
Temperature:Pathology.Results	0.999	2.000	≥0.25	ns
Richness
Temperature	8.625	2.000	0.013	*
Pathology.Results	0.840	1.000	≥0.25	ns
Temperature:Pathology.Results	0.495	2.000	≥0.25	ns
Phylogenetic
Temperature	8.874	2.000	0.012	*
Pathology.Results	0.625	1.000	≥0.25	ns
Temperature:Pathology.Results	0.674	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Pathology.Results	positive	negative	0.223	0.732	0.305	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	0.348	0.383	0.908	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.899	0.659	−1.365	0.172	Pathology.Results	35
Simpson
28	Alpha.Score	Pathology.Results	positive	negative	0.375	0.754	0.497	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	0.429	0.394	1.089	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.779	0.692	−1.125	≥0.25	Pathology.Results	35
Richness
28	Alpha.Score	Pathology.Results	positive	negative	−0.390	0.608	−0.641	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	−0.080	0.301	−0.267	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.611	0.552	−1.107	≥0.25	Pathology.Results	35
Phylogenetic
28	Alpha.Score	Pathology.Results	positive	negative	−0.390	0.577	−0.676	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	−0.017	0.280	−0.062	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.502	0.524	−0.958	≥0.25	Pathology.Results	35

Addl. Metrics

5B)

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature*Pathology.Results); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.569	0.001	***
Pathology.Results	1.000	0.263	0.013	1.471	0.179	ns
Temperature:Pathology.Results	2.000	0.404	0.019	1.127	≥0.25	ns
Residual	83.000	14.863	0.716	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.751	0.001	***
Pathology.Results	1.000	0.488	0.016	1.570	0.029	*
Temperature:Pathology.Results	2.000	0.679	0.022	1.093	≥0.25	ns
Residual	83.000	25.781	0.845	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.651	0.001	***
Pathology.Results	1.000	0.234	0.015	1.622	0.141	ns
Temperature:Pathology.Results	2.000	0.264	0.017	0.915	≥0.25	ns
Residual	83.000	11.980	0.756	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.Path	5.000	0.520	0.104	4.652	<0.001	***
Residual	83.000	1.855	0.022	NA	NA	NA
canberra
Temp.Path	5.000	0.625	0.125	13.830	<0.001	****
Residual	83.000	0.750	0.009	NA	NA	NA
gunifrac
Temp.Path	5.000	0.436	0.087	6.954	<0.001	****
Residual	83.000	1.040	0.013	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.Path	28°C__pos	28°C__neg	0.022	−0.165	0.209	≥0.25	ns
Distance	Temp.Path	32°C__neg	28°C__neg	−0.086	−0.277	0.105	≥0.25	ns
Distance	Temp.Path	32°C__pos	28°C__neg	0.015	−0.176	0.206	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__neg	−0.126	−0.298	0.046	≥0.25	ns
Distance	Temp.Path	35°C__pos	28°C__neg	−0.434	−0.896	0.029	0.079	ns
Distance	Temp.Path	32°C__neg	28°C__pos	−0.108	−0.263	0.046	≥0.25	ns
Distance	Temp.Path	32°C__pos	28°C__pos	−0.007	−0.161	0.148	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__pos	−0.148	−0.278	−0.017	0.017	*
Distance	Temp.Path	35°C__pos	28°C__pos	−0.456	−0.905	−0.007	0.044	*
Distance	Temp.Path	32°C__pos	32°C__neg	0.101	−0.058	0.261	≥0.25	ns
Distance	Temp.Path	35°C__neg	32°C__neg	−0.039	−0.175	0.096	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__neg	−0.348	−0.798	0.103	0.226	ns
Distance	Temp.Path	35°C__neg	32°C__pos	−0.141	−0.277	−0.005	0.038	*
Distance	Temp.Path	35°C__pos	32°C__pos	−0.449	−0.899	0.001	0.051	ns
Distance	Temp.Path	35°C__pos	35°C__neg	−0.308	−0.751	0.135	≥0.25	ns
canberra
Distance	Temp.Path	28°C__pos	28°C__neg	0.045	−0.074	0.164	≥0.25	ns
Distance	Temp.Path	32°C__neg	28°C__neg	−0.123	−0.244	−0.002	0.045	*
Distance	Temp.Path	32°C__pos	28°C__neg	−0.042	−0.164	0.079	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__neg	−0.105	−0.214	0.005	0.068	ns
Distance	Temp.Path	35°C__pos	28°C__neg	−0.588	−0.883	−0.294	<0.001	****
Distance	Temp.Path	32°C__neg	28°C__pos	−0.168	−0.267	−0.070	<0.001	****
Distance	Temp.Path	32°C__pos	28°C__pos	−0.088	−0.186	0.010	0.107	ns
Distance	Temp.Path	35°C__neg	28°C__pos	−0.150	−0.233	−0.067	<0.001	****
Distance	Temp.Path	35°C__pos	28°C__pos	−0.634	−0.919	−0.348	<0.001	****
Distance	Temp.Path	32°C__pos	32°C__neg	0.080	−0.021	0.182	0.199	ns
Distance	Temp.Path	35°C__neg	32°C__neg	0.018	−0.068	0.105	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__neg	−0.465	−0.752	−0.179	<0.001	***
Distance	Temp.Path	35°C__neg	32°C__pos	−0.062	−0.149	0.024	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__pos	−0.546	−0.832	−0.259	<0.001	****
Distance	Temp.Path	35°C__pos	35°C__neg	−0.484	−0.765	−0.202	<0.001	****
gunifrac
Distance	Temp.Path	28°C__pos	28°C__neg	0.011	−0.129	0.151	≥0.25	ns
Distance	Temp.Path	32°C__neg	28°C__neg	−0.091	−0.234	0.052	≥0.25	ns
Distance	Temp.Path	32°C__pos	28°C__neg	−0.014	−0.157	0.129	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__neg	−0.129	−0.258	0.000	0.049	*
Distance	Temp.Path	35°C__pos	28°C__neg	−0.411	−0.758	−0.065	0.011	*
Distance	Temp.Path	32°C__neg	28°C__pos	−0.102	−0.218	0.014	0.115	ns
Distance	Temp.Path	32°C__pos	28°C__pos	−0.025	−0.141	0.091	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__pos	−0.140	−0.238	−0.043	<0.001	***
Distance	Temp.Path	35°C__pos	28°C__pos	−0.423	−0.759	−0.087	0.006	**
Distance	Temp.Path	32°C__pos	32°C__neg	0.077	−0.042	0.196	≥0.25	ns
Distance	Temp.Path	35°C__neg	32°C__neg	−0.038	−0.140	0.064	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__neg	−0.321	−0.658	0.017	0.072	ns
Distance	Temp.Path	35°C__neg	32°C__pos	−0.115	−0.217	−0.013	0.017	*
Distance	Temp.Path	35°C__pos	32°C__pos	−0.398	−0.735	−0.060	0.011	*
Distance	Temp.Path	35°C__pos	35°C__neg	−0.283	−0.614	0.049	0.140	ns

Addl. Metrics

5C)

Tables

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GLM

GLM Results
glm(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	0.000	0.778	0.000	≥0.25	ns
Alpha.Score	0.000	1.808	0.000	≥0.25	ns
ClusterLow	6.524	1.067	6.115	<0.001	****
ClusterHigh	−2.610	3.512	−0.743	≥0.25	ns
Alpha.Score:ClusterLow	−10.090	4.999	−2.018	0.047	*
Alpha.Score:ClusterHigh	13.706	4.766	2.876	0.005	**
Simpson
(Intercept)	0.000	0.741	0.000	≥0.25	ns
Alpha.Score	0.000	1.605	0.000	≥0.25	ns
ClusterLow	6.842	1.041	6.573	<0.001	****
ClusterHigh	−3.247	3.900	−0.833	≥0.25	ns
Alpha.Score:ClusterLow	−12.165	4.647	−2.618	0.011	*
Alpha.Score:ClusterHigh	14.198	5.090	2.789	0.007	**
Richness
(Intercept)	0.000	0.914	0.000	≥0.25	ns
Alpha.Score	0.000	1.983	0.000	≥0.25	ns
ClusterLow	6.105	1.294	4.717	<0.001	****
ClusterHigh	7.362	2.644	2.784	0.007	**
Alpha.Score:ClusterLow	−3.491	4.463	−0.782	≥0.25	ns
Alpha.Score:ClusterHigh	0.772	4.163	0.185	≥0.25	ns
Phylogenetic
(Intercept)	0.000	0.912	0.000	≥0.25	ns
Alpha.Score	0.000	2.061	0.000	≥0.25	ns
ClusterLow	6.230	1.346	4.629	<0.001	****
ClusterHigh	8.137	2.589	3.143	0.002	**
Alpha.Score:ClusterLow	−3.872	4.662	−0.831	≥0.25	ns
Alpha.Score:ClusterHigh	−0.429	4.237	−0.101	≥0.25	ns

GLM Results
glm(Alpha.Score ~ Temperature*Total.Worm.Count); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−1.421	0.364	−3.908	<0.001	***
Temperature32	1.184	0.430	2.753	0.007	**
Temperature35	1.024	0.406	2.521	0.014	*
Total.Worm.Count	0.095	0.051	1.851	0.068	ns
Temperature32:Total.Worm.Count	−0.030	0.066	−0.453	≥0.25	ns
Temperature35:Total.Worm.Count	−0.994	1.255	−0.792	≥0.25	ns
Simpson
(Intercept)	−1.579	0.386	−4.092	<0.001	****
Temperature32	1.429	0.451	3.169	0.002	**
Temperature35	1.370	0.427	3.211	0.002	**
Total.Worm.Count	0.111	0.053	2.078	0.041	*
Temperature32:Total.Worm.Count	−0.047	0.068	−0.687	≥0.25	ns
Temperature35:Total.Worm.Count	−0.889	1.182	−0.752	≥0.25	ns
Richness
(Intercept)	−0.749	0.284	−2.641	0.010	**
Temperature32	0.726	0.349	2.078	0.041	*
Temperature35	0.326	0.327	0.998	≥0.25	ns
Total.Worm.Count	0.014	0.043	0.329	≥0.25	ns
Temperature32:Total.Worm.Count	0.000	0.056	−0.006	≥0.25	ns
Temperature35:Total.Worm.Count	−0.625	1.049	−0.596	≥0.25	ns
Phylogenetic
(Intercept)	−0.705	0.267	−2.643	0.010	**
Temperature32	0.617	0.329	1.875	0.064	ns
Temperature35	0.199	0.308	0.645	≥0.25	ns
Total.Worm.Count	0.005	0.041	0.123	≥0.25	ns
Temperature32:Total.Worm.Count	0.012	0.053	0.221	≥0.25	ns
Temperature35:Total.Worm.Count	−0.507	0.986	−0.514	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Total.Worm.Count ~ Alpha.Score*Cluster), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Alpha.Score	0.143	1.000	≥0.25	ns
Cluster	162.212	2.000	<0.001	****
Alpha.Score:Cluster	14.204	2.000	<0.001	***
Simpson
Alpha.Score	0.002	1.000	≥0.25	ns
Cluster	168.513	2.000	<0.001	****
Alpha.Score:Cluster	16.419	2.000	<0.001	***
Richness
Alpha.Score	0.066	1.000	≥0.25	ns
Cluster	145.539	2.000	<0.001	****
Alpha.Score:Cluster	0.741	2.000	≥0.25	ns
Phylogenetic
Alpha.Score	0.175	1.000	≥0.25	ns
Cluster	145.520	2.000	<0.001	****
Alpha.Score:Cluster	0.696	2.000	≥0.25	ns

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	13.111	2.000	0.001	**
Total.Worm.Count	5.701	1.000	0.017	*
Temperature:Total.Worm.Count	0.911	2.000	≥0.25	ns
Simpson
Temperature	18.031	2.000	<0.001	***
Total.Worm.Count	6.203	1.000	0.013	*
Temperature:Total.Worm.Count	1.061	2.000	≥0.25	ns
Richness
Temperature	8.429	2.000	0.015	*
Total.Worm.Count	0.239	1.000	≥0.25	ns
Temperature:Total.Worm.Count	0.386	2.000	≥0.25	ns
Phylogenetic
Temperature	8.411	2.000	0.015	*
Total.Worm.Count	0.196	1.000	≥0.25	ns
Temperature:Total.Worm.Count	0.339	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Cluster	Low	Other	−1.594	0.642	−2.483	0.035	Cluster	28
28	Alpha.Score	Cluster	High	Other	2.844	0.672	4.233	<0.001	Cluster	28
28	Alpha.Score	Cluster	High	Low	4.438	0.773	5.741	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−1.391	0.495	−2.808	0.013	Cluster	32
32	Alpha.Score	Cluster	High	Other	1.147	0.333	3.443	0.002	Cluster	32
32	Alpha.Score	Cluster	High	Low	2.538	0.528	4.811	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.899	0.659	−1.365	0.172	Cluster	35
Simpson
28	Alpha.Score	Cluster	Low	Other	−1.487	0.656	−2.266	0.060	Cluster	28
28	Alpha.Score	Cluster	High	Other	2.970	0.665	4.466	<0.001	Cluster	28
28	Alpha.Score	Cluster	High	Low	4.457	0.767	5.810	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−1.233	0.487	−2.531	0.030	Cluster	32
32	Alpha.Score	Cluster	High	Other	1.264	0.354	3.573	<0.001	Cluster	32
32	Alpha.Score	Cluster	High	Low	2.497	0.528	4.729	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.779	0.692	−1.125	≥0.25	Cluster	35
Richness
28	Alpha.Score	Cluster	Low	Other	−1.430	0.589	−2.429	0.040	Cluster	28
28	Alpha.Score	Cluster	High	Other	1.367	0.653	2.095	0.090	Cluster	28
28	Alpha.Score	Cluster	High	Low	2.797	0.692	4.043	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−1.183	0.387	−3.052	0.006	Cluster	32
32	Alpha.Score	Cluster	High	Other	0.429	0.278	1.543	≥0.25	Cluster	32
32	Alpha.Score	Cluster	High	Low	1.611	0.410	3.934	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.611	0.552	−1.107	≥0.25	Cluster	35
Phylogenetic
28	Alpha.Score	Cluster	Low	Other	−1.315	0.550	−2.389	0.044	Cluster	28
28	Alpha.Score	Cluster	High	Other	1.204	0.611	1.970	0.119	Cluster	28
28	Alpha.Score	Cluster	High	Low	2.519	0.638	3.946	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−0.994	0.360	−2.760	0.015	Cluster	32
32	Alpha.Score	Cluster	High	Other	0.437	0.262	1.667	0.214	Cluster	32
32	Alpha.Score	Cluster	High	Low	1.431	0.381	3.760	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.502	0.524	−0.958	≥0.25	Cluster	35

Addl. Metrics

5D)

Tables

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature*Cluster); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	18.730	0.001	***
Cluster	2.000	3.594	0.173	12.902	0.001	***
Temperature:Cluster	3.000	0.653	0.031	1.564	0.097	ns
Residual	81.000	11.282	0.544	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	6.064	0.001	***
Cluster	2.000	1.928	0.063	3.273	0.001	***
Temperature:Cluster	3.000	1.157	0.038	1.310	0.052	ns
Residual	81.000	23.862	0.782	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	14.844	0.001	***
Cluster	2.000	2.657	0.168	11.728	0.001	***
Temperature:Cluster	3.000	0.645	0.041	1.898	0.027	*
Residual	81.000	9.176	0.579	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

ADONIS2
adonis2(Beta Distance ~ Temperature*Total.Worm.Count); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.815	0.001	***
Total.Worm.Count	1.000	0.588	0.028	3.341	0.009	**
Temperature:Total.Worm.Count	2.000	0.326	0.016	0.925	≥0.25	ns
Residual	83.000	14.616	0.704	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.754	0.001	***
Total.Worm.Count	1.000	0.498	0.016	1.605	0.032	*
Temperature:Total.Worm.Count	2.000	0.678	0.022	1.093	≥0.25	ns
Residual	83.000	25.771	0.844	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.736	0.001	***
Total.Worm.Count	1.000	0.350	0.022	2.443	0.031	*
Temperature:Total.Worm.Count	2.000	0.235	0.015	0.822	≥0.25	ns
Residual	83.000	11.892	0.751	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature*Cluster); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Cluster	2.000	0.019	0.009	0.383	≥0.25	ns
Residual	86.000	2.116	0.025	NA	NA	NA
canberra
Cluster	2.000	0.167	0.084	10.322	<0.001	****
Residual	86.000	0.697	0.008	NA	NA	NA
gunifrac
Cluster	2.000	0.016	0.008	0.628	≥0.25	ns
Residual	86.000	1.078	0.013	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Cluster); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Cluster	Low	Other	−0.037	−0.138	0.064	≥0.25	ns
Distance	Cluster	High	Other	−0.005	−0.114	0.104	≥0.25	ns
Distance	Cluster	High	Low	0.032	−0.099	0.163	≥0.25	ns
canberra
Distance	Cluster	Low	Other	0.098	0.040	0.156	<0.001	***
Distance	Cluster	High	Other	−0.031	−0.093	0.032	≥0.25	ns
Distance	Cluster	High	Low	−0.129	−0.204	−0.053	<0.001	***
gunifrac
Distance	Cluster	Low	Other	0.017	−0.055	0.089	≥0.25	ns
Distance	Cluster	High	Other	−0.027	−0.104	0.051	≥0.25	ns
Distance	Cluster	High	Low	−0.044	−0.137	0.050	≥0.25	ns

Addl. Metrics

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S5A

S5B

S5C

S5D

Click on tabs to cycle through different cluster comparisons between alpha and beta diversity.

Phylogenetic

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric	term	df	SumOfSqs	R2	statistic	p.value	p.adj.sig
Phylogenetic
bray	Cluster	2.000	5.369	0.259	15.012	0.001	**
bray	Residual	86.000	15.379	0.741	NA	NA	NA
bray	Total	88.000	20.748	1.000	NA	NA	NA
canberra	Cluster	2.000	3.204	0.105	5.043	0.001	**
canberra	Residual	86.000	27.317	0.895	NA	NA	NA
canberra	Total	88.000	30.520	1.000	NA	NA	NA
gunifrac	Cluster	2.000	3.692	0.233	13.068	0.001	**
gunifrac	Residual	86.000	12.149	0.767	NA	NA	NA
gunifrac	Total	88.000	15.841	1.000	NA	NA	NA

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric	term	df	sumsq	meansq	statistic	p.value	p.adj.sig
Phylogenetic
bray	Groups	2.000	0.039	0.019	0.754	≥0.25	ns
bray	Residuals	86.000	2.218	0.026	NA	NA	NA
canberra	Groups	2.000	0.221	0.110	14.247	<0.001	****
canberra	Residuals	86.000	0.666	0.008	NA	NA	NA
gunifrac	Groups	2.000	0.072	0.036	2.725	0.071	ns
gunifrac	Residuals	86.000	1.131	0.013	NA	NA	NA

Richness

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric	term	df	SumOfSqs	R2	statistic	p.value	p.adj.sig
Richness
bray	Cluster	2.000	5.363	0.258	14.990	0.001	**
bray	Residual	86.000	15.385	0.742	NA	NA	NA
bray	Total	88.000	20.748	1.000	NA	NA	NA
canberra	Cluster	2.000	3.356	0.110	5.312	0.001	**
canberra	Residual	86.000	27.165	0.890	NA	NA	NA
canberra	Total	88.000	30.520	1.000	NA	NA	NA
gunifrac	Cluster	2.000	3.835	0.242	13.736	0.001	**
gunifrac	Residual	86.000	12.006	0.758	NA	NA	NA
gunifrac	Total	88.000	15.841	1.000	NA	NA	NA

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric	term	df	sumsq	meansq	statistic	p.value	p.adj.sig
Richness
bray	Groups	2.000	0.028	0.014	0.535	≥0.25	ns
bray	Residuals	86.000	2.263	0.026	NA	NA	NA
canberra	Groups	2.000	0.216	0.108	13.957	<0.001	****
canberra	Residuals	86.000	0.664	0.008	NA	NA	NA
gunifrac	Groups	2.000	0.050	0.025	1.873	0.160	ns
gunifrac	Residuals	86.000	1.155	0.013	NA	NA	NA

Shannon

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric	term	df	SumOfSqs	R2	statistic	p.value	p.adj.sig
Shannon
bray	Cluster	2.000	5.943	0.286	17.260	0.001	**
bray	Residual	86.000	14.805	0.714	NA	NA	NA
bray	Total	88.000	20.748	1.000	NA	NA	NA
canberra	Cluster	2.000	3.188	0.104	5.016	0.001	**
canberra	Residual	86.000	27.332	0.896	NA	NA	NA
canberra	Total	88.000	30.520	1.000	NA	NA	NA
gunifrac	Cluster	2.000	4.229	0.267	15.660	0.001	**
gunifrac	Residual	86.000	11.612	0.733	NA	NA	NA
gunifrac	Total	88.000	15.841	1.000	NA	NA	NA

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric	term	df	sumsq	meansq	statistic	p.value	p.adj.sig
Shannon
bray	Groups	2.000	0.019	0.009	0.383	≥0.25	ns
bray	Residuals	86.000	2.116	0.025	NA	NA	NA
canberra	Groups	2.000	0.167	0.084	10.322	<0.001	****
canberra	Residuals	86.000	0.697	0.008	NA	NA	NA
gunifrac	Groups	2.000	0.016	0.008	0.628	≥0.25	ns
gunifrac	Residuals	86.000	1.078	0.013	NA	NA	NA

Simpsons

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric	term	df	SumOfSqs	R2	statistic	p.value	p.adj.sig
Simpson
bray	Cluster	2.000	5.943	0.286	17.260	0.001	**
bray	Residual	86.000	14.805	0.714	NA	NA	NA
bray	Total	88.000	20.748	1.000	NA	NA	NA
canberra	Cluster	2.000	3.188	0.104	5.016	0.001	**
canberra	Residual	86.000	27.332	0.896	NA	NA	NA
canberra	Total	88.000	30.520	1.000	NA	NA	NA
gunifrac	Cluster	2.000	4.229	0.267	15.660	0.001	**
gunifrac	Residual	86.000	11.612	0.733	NA	NA	NA
gunifrac	Total	88.000	15.841	1.000	NA	NA	NA

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric	term	df	sumsq	meansq	statistic	p.value	p.adj.sig
Simpson
bray	Groups	2.000	0.019	0.009	0.383	≥0.25	ns
bray	Residuals	86.000	2.116	0.025	NA	NA	NA
canberra	Groups	2.000	0.167	0.084	10.322	<0.001	****
canberra	Residuals	86.000	0.697	0.008	NA	NA	NA
gunifrac	Groups	2.000	0.016	0.008	0.628	≥0.25	ns
gunifrac	Residuals	86.000	1.078	0.013	NA	NA	NA

Cluster Counts

Counts of samples per cluster and temperature by alpha-diversity metric. Click on tabs to reveal tables.

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Simpson

Temperature	Count	Percentage
Simpson - High
28	5	33.33
32	10	66.67
Simpson - Low
28	12	66.67
32	5	27.78
35	1	5.56
Simpson - Other
28	8	14.29
32	15	26.79
35	33	58.93

Shannon

Temperature	Count	Percentage
Shannon - High
28	5	33.33
32	10	66.67
Shannon - Low
28	12	66.67
32	5	27.78
35	1	5.56
Shannon - Other
28	8	14.29
32	15	26.79
35	33	58.93

Richness

Temperature	Count	Percentage
Richness - High
28	5	35.71
32	9	64.29
Richness - Low
28	12	63.16
32	6	31.58
35	1	5.26
Richness - Other
28	8	14.29
32	15	26.79
35	33	58.93

Phylogenetic

Temperature	Count	Percentage
Phylogenetic - High
28	4	30.77
32	9	69.23
Phylogenetic - Low
28	13	65.00
32	6	30.00
35	1	5.00
Phylogenetic - Other
28	8	14.29
32	15	26.79
35	33	58.93

Fig. 6) Parasite exposure exacerbates water temperature differences in gut microbiome structure

Fig. 6 Comparison of the effects of water temperature on the gut microbiome between parasite exposed fish and parasite unexposed fish. (A) Simpson’s Index for diversity of parasite unexposed and pre-exposed fish at 0 days post exposure (dpe). Prior to parasite exposure gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature. (B) Capscale ordinations based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main and interaction effects of temperature and parasite exposure (treatment) of pre-exposure samples at 0 dpe. (C) Simpson’s Index for diversity of parasite unexposed and exposed fish. Gut microbial alpha-diversity significantly differs between parasite exposed fish reared at 28°C and 32°C water temperature relative to unexposed control fish, but gut microbial alpha-diversity does not differ between parasite unexposed and exposed fish reared at 35°C water temperature. (D) Capscale ordinations based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main and interaction effects of temperature and parasite exposure (treatment) of post-exposure samples after 0 dpe. The analysis shows gut microbiome composition differs between fish reared at different water temperatures prior to parasite exposure, and parasite exposure further drives these temperature associated differences in microbiome community composition. Ribbons and ellipses indicate 95% confidence interval. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

6A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.218	0.269	−0.811	≥0.25	ns
Temperature32	0.345	0.379	0.910	≥0.25	ns
Temperature35	0.693	0.384	1.803	0.077	ns
TreatmentExposed	0.274	0.379	0.723	≥0.25	ns
Temperature32:TreatmentExposed	0.311	0.544	0.572	≥0.25	ns
Temperature35:TreatmentExposed	0.041	0.550	0.075	≥0.25	ns
Simpson
(Intercept)	−0.241	0.252	−0.958	≥0.25	ns
Temperature32	0.352	0.355	0.990	≥0.25	ns
Temperature35	0.961	0.367	2.619	0.011	*
TreatmentExposed	0.223	0.355	0.627	≥0.25	ns
Temperature32:TreatmentExposed	0.486	0.513	0.948	≥0.25	ns
Temperature35:TreatmentExposed	0.017	0.525	0.033	≥0.25	ns
Richness
(Intercept)	−0.145	0.270	−0.538	≥0.25	ns
Temperature32	0.550	0.385	1.430	0.158	ns
Temperature35	0.043	0.381	0.112	≥0.25	ns
TreatmentExposed	0.402	0.382	1.050	≥0.25	ns
Temperature32:TreatmentExposed	0.020	0.554	0.036	≥0.25	ns
Temperature35:TreatmentExposed	0.287	0.545	0.526	≥0.25	ns
Phylogenetic
(Intercept)	−0.253	0.254	−0.996	≥0.25	ns
Temperature32	0.553	0.360	1.536	0.130	ns
Temperature35	0.171	0.358	0.478	≥0.25	ns
TreatmentExposed	0.335	0.358	0.937	≥0.25	ns
Temperature32:TreatmentExposed	−0.018	0.512	−0.035	≥0.25	ns
Temperature35:TreatmentExposed	0.102	0.507	0.201	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Pre-Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	7.264	2.000	0.026	*
Treatment	3.044	1.000	0.081	ns
Temperature:Treatment	0.380	2.000	≥0.25	ns
Simpson
Temperature	14.462	2.000	<0.001	***
Treatment	3.406	1.000	0.065	ns
Temperature:Treatment	1.122	2.000	≥0.25	ns
Richness
Temperature	4.311	2.000	0.116	ns
Treatment	5.067	1.000	0.024	*
Temperature:Treatment	0.340	2.000	≥0.25	ns
Phylogenetic
Temperature	4.600	2.000	0.100	ns
Treatment	3.056	1.000	0.080	ns
Temperature:Treatment	0.064	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Treatment	Exposed	Control	0.274	0.364	0.753	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.585	0.451	1.298	0.194	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.315	0.344	0.917	≥0.25	Treatment	35
Simpson
28	Alpha.Score	Treatment	Exposed	Control	0.223	0.337	0.662	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.709	0.426	1.663	0.096	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.240	0.339	0.708	≥0.25	Treatment	35
Richness
28	Alpha.Score	Treatment	Exposed	Control	0.402	0.361	1.112	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.422	0.411	1.027	≥0.25	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.689	0.400	1.721	0.085	Treatment	35
Phylogenetic
28	Alpha.Score	Treatment	Exposed	Control	0.335	0.344	0.974	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.317	0.348	0.911	≥0.25	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.437	0.389	1.123	≥0.25	Treatment	35

Addl. Metrics

6B)

Tables

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Pre-Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	0.871	0.188	7.178	0.001	***
Treatment	1.000	0.244	0.053	4.025	0.009	**
Temperature:Treatment	2.000	0.245	0.053	2.019	0.047	*
Residual	54.000	3.275	0.707	NA	NA	NA
Total	59.000	4.635	1.000	NA	NA	NA
canberra
Temperature	2.000	1.652	0.132	4.615	0.001	***
Treatment	1.000	0.533	0.043	2.979	0.001	***
Temperature:Treatment	2.000	0.667	0.053	1.864	0.001	***
Residual	54.000	9.665	0.772	NA	NA	NA
Total	59.000	12.517	1.000	NA	NA	NA
gunifrac
Temperature	2.000	0.647	0.153	5.604	0.001	***
Treatment	1.000	0.288	0.068	4.987	0.003	**
Temperature:Treatment	2.000	0.177	0.042	1.537	0.110	ns
Residual	54.000	3.117	0.737	NA	NA	NA
Total	59.000	4.229	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Pre-Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.Treat	5.000	0.103	0.021	2.738	0.028	*
Residual	54.000	0.405	0.007	NA	NA	NA
canberra
Temp.Treat	5.000	0.014	0.003	0.453	≥0.25	ns
Residual	54.000	0.329	0.006	NA	NA	NA
gunifrac
Temp.Treat	5.000	0.054	0.011	1.564	0.186	ns
Residual	54.000	0.369	0.007	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Pre-Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.027	−0.088	0.141	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Control	0.095	−0.019	0.210	0.153	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.110	−0.004	0.225	0.065	ns
Distance	Temp.Treat	35°C__Control	28°C__Control	0.015	−0.099	0.130	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.069	−0.045	0.183	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	0.069	−0.046	0.183	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	0.084	−0.031	0.198	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.011	−0.126	0.103	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	0.042	−0.072	0.157	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.015	−0.100	0.129	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Control	−0.080	−0.195	0.034	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	−0.026	−0.141	0.088	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.095	−0.209	0.019	0.156	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.041	−0.156	0.073	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.054	−0.061	0.168	≥0.25	ns
canberra
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.011	−0.092	0.114	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Control	0.035	−0.068	0.138	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.044	−0.059	0.147	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Control	0.010	−0.094	0.113	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.016	−0.087	0.119	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	0.024	−0.079	0.127	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	0.033	−0.070	0.136	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.001	−0.104	0.102	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	0.005	−0.098	0.108	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.009	−0.094	0.112	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Control	−0.025	−0.128	0.078	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	−0.019	−0.122	0.084	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.034	−0.137	0.069	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.028	−0.131	0.075	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.006	−0.097	0.109	≥0.25	ns
gunifrac
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.003	−0.106	0.112	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Control	0.051	−0.058	0.160	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.078	−0.032	0.187	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Control	0.000	−0.109	0.110	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.042	−0.067	0.151	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	0.048	−0.061	0.158	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	0.075	−0.035	0.184	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.002	−0.112	0.107	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	0.039	−0.070	0.148	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.026	−0.083	0.136	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Control	−0.051	−0.160	0.059	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	−0.009	−0.118	0.100	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.077	−0.186	0.032	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.036	−0.145	0.074	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.041	−0.068	0.151	≥0.25	ns

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6C)

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.441	0.126	−3.499	<0.001	***
Temperature32	−0.064	0.177	−0.364	≥0.25	ns
Temperature35	−0.087	0.182	−0.476	≥0.25	ns
TreatmentExposed	−0.539	0.210	−2.562	0.011	*
Temperature32:TreatmentExposed	0.953	0.280	3.405	<0.001	***
Temperature35:TreatmentExposed	0.561	0.281	1.996	0.047	*
Simpson
(Intercept)	−0.299	0.133	−2.252	0.025	*
Temperature32	−0.059	0.186	−0.319	≥0.25	ns
Temperature35	0.080	0.190	0.419	≥0.25	ns
TreatmentExposed	−0.724	0.225	−3.223	0.001	**
Temperature32:TreatmentExposed	1.106	0.298	3.715	<0.001	***
Temperature35:TreatmentExposed	0.673	0.297	2.270	0.024	*
Richness
(Intercept)	−0.206	0.101	−2.040	0.043	*
Temperature32	−0.048	0.141	−0.343	≥0.25	ns
Temperature35	−0.317	0.147	−2.164	0.032	*
TreatmentExposed	−0.228	0.160	−1.421	0.157	ns
Temperature32:TreatmentExposed	0.829	0.219	3.776	<0.001	***
Temperature35:TreatmentExposed	0.691	0.219	3.155	0.002	**
Phylogenetic
(Intercept)	−0.143	0.095	−1.507	0.134	ns
Temperature32	−0.035	0.132	−0.268	≥0.25	ns
Temperature35	−0.319	0.138	−2.320	0.021	*
TreatmentExposed	−0.192	0.150	−1.280	0.202	ns
Temperature32:TreatmentExposed	0.780	0.206	3.783	<0.001	***
Temperature35:TreatmentExposed	0.639	0.205	3.111	0.002	**

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Post-Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	5.908	2.000	0.052	ns
Treatment	0.001	1.000	≥0.25	ns
Temperature:Treatment	11.784	2.000	0.003	**
Simpson
Temperature	8.400	2.000	0.015	*
Treatment	0.547	1.000	≥0.25	ns
Temperature:Treatment	14.128	2.000	<0.001	***
Richness
Temperature	11.325	2.000	0.003	**
Treatment	11.824	1.000	<0.001	***
Temperature:Treatment	16.178	2.000	<0.001	***
Phylogenetic
Temperature	13.345	2.000	0.001	**
Treatment	13.294	1.000	<0.001	***
Temperature:Treatment	16.080	2.000	<0.001	***

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Treatment	Exposed	Control	−0.539	0.280	−1.924	0.054	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.414	0.180	2.298	0.022	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.022	0.111	0.200	≥0.25	Treatment	35
Simpson
28	Alpha.Score	Treatment	Exposed	Control	−0.724	0.290	−2.492	0.013	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.382	0.190	2.013	0.044	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	−0.051	0.130	−0.387	≥0.25	Treatment	35
Richness
28	Alpha.Score	Treatment	Exposed	Control	−0.228	0.223	−1.021	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.601	0.132	4.546	<0.001	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.463	0.092	5.021	<0.001	Treatment	35
Phylogenetic
28	Alpha.Score	Treatment	Exposed	Control	−0.192	0.210	−0.916	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.588	0.121	4.858	<0.001	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.447	0.090	4.961	<0.001	Treatment	35

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6D)

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ADONIS2

ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Post-Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	4.123	0.108	17.035	0.001	***
Treatment	1.000	7.411	0.194	61.235	0.001	***
Temperature:Treatment	2.000	3.168	0.083	13.088	0.001	***
Residual	194.000	23.478	0.615	NA	NA	NA
Total	199.000	38.179	1.000	NA	NA	NA
canberra
Temperature	2.000	4.561	0.074	9.502	0.001	***
Treatment	1.000	7.402	0.120	30.840	0.001	***
Temperature:Treatment	2.000	3.014	0.049	6.279	0.001	***
Residual	194.000	46.561	0.757	NA	NA	NA
Total	199.000	61.538	1.000	NA	NA	NA
gunifrac
Temperature	2.000	2.273	0.082	11.945	0.001	***
Treatment	1.000	4.718	0.170	49.577	0.001	***
Temperature:Treatment	2.000	2.273	0.082	11.944	0.001	***
Residual	194.000	18.460	0.666	NA	NA	NA
Total	199.000	27.724	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Post-Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.Treat	5.000	1.344	0.269	17.882	<0.001	****
Residual	194.000	2.917	0.015	NA	NA	NA
canberra
Temp.Treat	5.000	1.094	0.219	31.758	<0.001	****
Residual	194.000	1.337	0.007	NA	NA	NA
gunifrac
Temp.Treat	5.000	1.225	0.245	29.264	<0.001	****
Residual	194.000	1.624	0.008	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Post-Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.211	0.119	0.302	<0.001	****
Distance	Temp.Treat	32°C__Control	28°C__Control	−0.015	−0.096	0.066	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.168	0.081	0.255	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Control	0.013	−0.070	0.096	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.060	−0.024	0.144	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	−0.225	−0.316	−0.135	<0.001	****
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	−0.043	−0.138	0.053	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.197	−0.290	−0.105	<0.001	****
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	−0.150	−0.243	−0.057	<0.001	****
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.183	0.097	0.268	<0.001	****
Distance	Temp.Treat	35°C__Control	32°C__Control	0.028	−0.054	0.110	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	0.075	−0.008	0.158	0.100	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.155	−0.243	−0.067	<0.001	****
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.108	−0.196	−0.019	0.007	**
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.047	−0.038	0.132	≥0.25	ns
canberra
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.217	0.155	0.279	<0.001	****
Distance	Temp.Treat	32°C__Control	28°C__Control	−0.011	−0.066	0.044	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.107	0.048	0.166	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Control	0.013	−0.044	0.069	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.073	0.016	0.130	0.004	**
Distance	Temp.Treat	32°C__Control	28°C__Exposed	−0.228	−0.289	−0.167	<0.001	****
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	−0.110	−0.175	−0.045	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.204	−0.267	−0.142	<0.001	****
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	−0.144	−0.207	−0.081	<0.001	****
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.118	0.060	0.176	<0.001	****
Distance	Temp.Treat	35°C__Control	32°C__Control	0.024	−0.032	0.080	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	0.084	0.028	0.140	<0.001	***
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.094	−0.154	−0.035	<0.001	***
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.034	−0.094	0.026	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.060	0.003	0.118	0.035	*
gunifrac
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.197	0.129	0.266	<0.001	****
Distance	Temp.Treat	32°C__Control	28°C__Control	−0.023	−0.083	0.037	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.141	0.077	0.206	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Control	−0.011	−0.073	0.051	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.054	−0.009	0.117	0.134	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	−0.220	−0.288	−0.153	<0.001	****
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	−0.056	−0.128	0.015	0.211	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.209	−0.278	−0.140	<0.001	****
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	−0.143	−0.213	−0.074	<0.001	****
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.164	0.100	0.228	<0.001	****
Distance	Temp.Treat	35°C__Control	32°C__Control	0.012	−0.050	0.073	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	0.077	0.015	0.139	0.006	**
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.152	−0.218	−0.087	<0.001	****
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.087	−0.153	−0.021	0.003	**
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.065	0.002	0.129	0.039	*

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S6A

S6B

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ADONIS2

Temperature	term	df	SumOfSqs	R2	statistic	p.value	p.adj.sig
bray
28	Treatment	1.000	0.065	0.082	1.615	0.176	ns
28	Residual	18.000	0.723	0.918	NA	NA	NA
28	Total	19.000	0.788	1.000	NA	NA	NA
32	Treatment	1.000	0.195	0.109	2.191	0.087	ns
32	Residual	18.000	1.603	0.891	NA	NA	NA
32	Total	19.000	1.799	1.000	NA	NA	NA
35	Treatment	1.000	0.229	0.194	4.346	0.008	**
35	Residual	18.000	0.948	0.806	NA	NA	NA
35	Total	19.000	1.177	1.000	NA	NA	NA
canberra
28	Treatment	1.000	0.358	0.105	2.102	0.010	*
28	Residual	18.000	3.068	0.895	NA	NA	NA
28	Total	19.000	3.426	1.000	NA	NA	NA
32	Treatment	1.000	0.422	0.107	2.153	0.009	**
32	Residual	18.000	3.524	0.893	NA	NA	NA
32	Total	19.000	3.946	1.000	NA	NA	NA
35	Treatment	1.000	0.421	0.120	2.464	0.007	**
35	Residual	18.000	3.073	0.880	NA	NA	NA
35	Total	19.000	3.493	1.000	NA	NA	NA
gunifrac
28	Treatment	1.000	0.086	0.092	1.828	0.130	ns
28	Residual	18.000	0.845	0.908	NA	NA	NA
28	Total	19.000	0.931	1.000	NA	NA	NA
32	Treatment	1.000	0.174	0.113	2.296	0.062	ns
32	Residual	18.000	1.361	0.887	NA	NA	NA
32	Total	19.000	1.535	1.000	NA	NA	NA
35	Treatment	1.000	0.206	0.184	4.068	0.012	*
35	Residual	18.000	0.911	0.816	NA	NA	NA
35	Total	19.000	1.117	1.000	NA	NA	NA

Dispersion (ANOVA)

Temperature	term	df	sumsq	meansq	statistic	p.value	p.adj.sig
bray
28	Groups	1.000	0.004	0.004	0.609	0.445	ns
28	Residuals	18.000	0.105	0.006	NA	NA	NA
32	Groups	1.000	0.001	0.001	0.100	0.755	ns
32	Residuals	18.000	0.199	0.011	NA	NA	NA
35	Groups	1.000	0.014	0.014	2.589	0.125	ns
35	Residuals	18.000	0.101	0.006	NA	NA	NA
canberra
28	Groups	1.000	0.001	0.001	0.061	0.807	ns
28	Residuals	18.000	0.172	0.010	NA	NA	NA
32	Groups	1.000	0.000	0.000	0.078	0.783	ns
32	Residuals	18.000	0.092	0.005	NA	NA	NA
35	Groups	1.000	0.000	0.000	0.055	0.817	ns
35	Residuals	18.000	0.065	0.004	NA	NA	NA
gunifrac
28	Groups	1.000	0.000	0.000	0.004	0.948	ns
28	Residuals	18.000	0.165	0.009	NA	NA	NA
32	Groups	1.000	0.003	0.003	0.465	0.504	ns
32	Residuals	18.000	0.135	0.008	NA	NA	NA
35	Groups	1.000	0.009	0.009	2.238	0.152	ns
35	Residuals	18.000	0.069	0.004	NA	NA	NA

S6C

S6D

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ADONIS2

Temperature	term	df	SumOfSqs	R2	statistic	p.value	p.adj.sig
bray
28	Treatment	1.000	6.719	0.447	48.541	0.001	**
28	Residual	60.000	8.305	0.553	NA	NA	NA
28	Total	61.000	15.023	1.000	NA	NA	NA
32	Treatment	1.000	2.842	0.261	23.668	0.001	**
32	Residual	67.000	8.044	0.739	NA	NA	NA
32	Total	68.000	10.886	1.000	NA	NA	NA
35	Treatment	1.000	1.018	0.125	9.568	0.001	**
35	Residual	67.000	7.129	0.875	NA	NA	NA
35	Total	68.000	8.147	1.000	NA	NA	NA
canberra
28	Treatment	1.000	3.887	0.191	14.144	0.001	**
28	Residual	60.000	16.490	0.809	NA	NA	NA
28	Total	61.000	20.377	1.000	NA	NA	NA
32	Treatment	1.000	3.542	0.191	15.809	0.001	**
32	Residual	67.000	15.012	0.809	NA	NA	NA
32	Total	68.000	18.554	1.000	NA	NA	NA
35	Treatment	1.000	2.987	0.165	13.288	0.001	**
35	Residual	67.000	15.059	0.835	NA	NA	NA
35	Total	68.000	18.046	1.000	NA	NA	NA
gunifrac
28	Treatment	1.000	4.085	0.374	35.857	0.001	**
28	Residual	60.000	6.836	0.626	NA	NA	NA
28	Total	61.000	10.922	1.000	NA	NA	NA
32	Treatment	1.000	2.031	0.230	19.988	0.001	**
32	Residual	67.000	6.806	0.770	NA	NA	NA
32	Total	68.000	8.837	1.000	NA	NA	NA
35	Treatment	1.000	1.080	0.160	12.720	0.001	**
35	Residual	67.000	5.691	0.840	NA	NA	NA
35	Total	68.000	6.771	1.000	NA	NA	NA

Dispersion (ANOVA)

Temperature	term	df	sumsq	meansq	statistic	p.value	p.adj.sig
bray
28	Groups	1.000	0.662	0.662	53.570	0.000	****
28	Residuals	60.000	0.741	0.012	NA	NA	NA
32	Groups	1.000	0.567	0.567	54.435	0.000	****
32	Residuals	67.000	0.697	0.010	NA	NA	NA
35	Groups	1.000	0.038	0.038	1.735	0.192	ns
35	Residuals	67.000	1.479	0.022	NA	NA	NA
canberra
28	Groups	1.000	0.703	0.703	224.734	0.000	****
28	Residuals	60.000	0.188	0.003	NA	NA	NA
32	Groups	1.000	0.237	0.237	30.033	0.000	****
32	Residuals	67.000	0.530	0.008	NA	NA	NA
35	Groups	1.000	0.062	0.062	6.749	0.012	*
35	Residuals	67.000	0.620	0.009	NA	NA	NA
gunifrac
28	Groups	1.000	0.582	0.582	124.383	0.000	****
28	Residuals	60.000	0.281	0.005	NA	NA	NA
32	Groups	1.000	0.482	0.482	57.322	0.000	****
32	Residuals	67.000	0.564	0.008	NA	NA	NA
35	Groups	1.000	0.078	0.078	5.842	0.018	*
35	Residuals	67.000	0.896	0.013	NA	NA	NA

Fig. 7) Gut microbial abundance is significantly associated with environmental conditions and stressors

7) Gut microbial abundance is significantly associated with environmental conditions and stressors

Fig. 7 A heatmap of model coefficient values of the top 50 statistically significant abundant gut microbial taxa identified by MaAsLin2. The color of each cell represents the coefficient value and direction (red is positive, blue is negative). A “+” or “-” indicates a statistically significant association was observed between taxon abundance and a covariate. Gray colored cells indicate a significant effect was not observed.

Taxon	Temp: 32°C	Temp: 35°C	Time (DPE)	Cluster: Low	Parasite exposed	Infection present	Infection burden
Actinobacteriota
IMCC26207	+		-	-	-
Nocardiaceae Genus	+	+	+
PeM15 Genus	+		-	-	-
Bacteroidota
Barnesiellaceae Genus			-		-
Cloacibacterium		+	-
Flavobacterium	-	-	-	-	+
Fluviicola		-	-		-	-
Microscillaceae Genus	+	+	+	-
Terrimonas				-		+	+
env OPS 17 Genus		-		-	-
Chloroflexi
JG30 KF CM45 Genus			+	-	+	+
Cyanobacteria
Candidatus Obscuribacter	-	-	+	-	+	+
Obscuribacteraceae Genus	+		+	-		+	-
Vampirovibrionaceae Genus			+	-	+	+
Desulfobacterota
Desulfobacterota Genus				-	+	+
Firmicutes
Clostridium sensu stricto 3			+	-	+	+
Gemmatimonadota
Gemmatimonadaceae Genus		-		-	+	+
Myxococcota
Sandaracinaceae Genus	+	+	+	-	+	+
NB1-j
NB1 j Genus		+	+	-	+	+
Nitrospirota
Nitrospira		-	+	-	+	+
Patescibacteria
Saccharimonadales Genus	+	+	+	-	+
Planctomycetota
CL500 3			+	-	-	+
Gemmata		+	+	-	+	+
Pirellulaceae Genus	+	-	+	-	-
Planctomycetales Genus			+	-	+	+
Planctomycetes Genus			+	-	+	+
Rhodopirellula				-	+	+	-
SM1A02			+	-	+	+
Telmatocola			+	-	+	+
Proteobacteria
A0839 Genus	+			-	+	+
Alphaproteobacteria Genus		+	+	-	+	+
Bauldia	+		+	-	+	+
Beijerinckiaceae Genus		-	+	-		+	-
Bosea	-	-	-	-
Candidatus Alysiosphaera	+		+	-	+	+
Chitinibacter		+	+		+		-
Comamonadaceae Genus			-	-	+	+
Gammaproteobacteria Genus	-	-	+	-		+
Luteimonas	-			-		+	+
Methyloligellaceae Genus	+	+	+	-	+	+
Nordella		-		-	+	+
Plesiomonas	+	+			-	-	-
Pseudomonadales Genus	-	-	-	-	-	-
Rhizobacter				-	+	+	+
Rhizobiales Incertae Sedis Genus				-		+
Rhodobacteraceae Genus				-	+	+
Rhodovastum		-		-
Steroidobacteraceae Genus			+	-	+	+
Xanthobacteraceae Genus	-	-	+	-		+
Verrucomicrobiota
Oikopleura			+	-	+	+

Tables

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S7.1

Summary of Significant Associations
(per covariate)
Significance	Cluster (Low)	DPE (DPE)	Pathology.Results (positive)	Temperature (32)	Temperature (35)	Total.Worm.Count (Total.Worm.Count)	Treatment (Exposed)
Positive	1	73	117	42	37	49	74
Negative	192	36	5	47	54	13	35

S7.2

Summary of Significant Associations
(per taxonomic level)
Taxonomy	Count
Kingdom	3
Phylum	28
Class	48
Order	114
Family	183
Genus	277

Suppl.

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S7.1

All taxa with significant associations
MaAsLin2(log(qval)*sign(coef)); All fish
Taxon	Temp: 32°C	Temp: 35°C	Time (DPE)	Cluster: Low	Parasite exposed	Infection present	Infection burden
Acidobacteriota
Bryobacter							+
Holophagaceae Genus			-	-		+
JGI 0001001 H03				-		+	+
Paludibaculum Genus	+			-	+
Subgroup 17 Genus				-	+
Vicinamibacteraceae Genus				-		+	+
Actinobacteriota
Acidimicrobiia Genus		+			-
Agromyces		+			+
Brevibacterium			+
Conexibacter		+	+		+
Corynebacteriales Genus	-	-	+	-
Frankiales Genus			+	-	+	+
Gaiella				-			+
Gaiellales Genus	+		+	-
Glutamicibacter				-
Gordonia	+	+	+	-
IMCC26207	+		-	-	-
IMCC26256 Genus	+		+	-		+
Iamia		-	+	-
Ilumatobacteraceae Genus			+		+
Kineosporiaceae Genus			+
Leucobacter			-
Longivirga		+		-
Microtrichaceae Genus							+
Microtrichales Genus	+	+	-	-
Mycobacterium	-		+	-	-	+
Nakamurella				-
Nocardia				-		+
Nocardiaceae Genus	+	+	+
Nocardioides			+	-
PeM15 Genus	+		-	-	-
Rhodococcus	+	+	+		-
Smaragdicoccus	+	+	+
Thermoleophilia Genus				-	+		+
Armatimonadota
Armatimonadota Genus							+
Bacteroidota
AKYH767 Genus	-			-		+
Aurantisolimonas			-		-
Bacteroidia Genus	-		-	-		+
Barnesiellaceae Genus			-		-
Candidatus Amoebophilus			+
Chitinophagaceae Genus	-			-		+	+
Chitinophagales Genus	-			-		+
Chryseobacterium		+
Cloacibacterium		+	-
Cytophaga	-				+
Cytophagales Genus	-			-		+
Dinghuibacter	+	+		-	-
Ferruginibacter	-	-		-		+	+
Flavihumibacter		-	-	-	-
Flavobacterium	-	-	-	-	+
Fluviicola		-	-		-	-
Haliscomenobacter	-	-		-		+
Hassallia				-			+
Kapabacteriales Genus	-			-		+
Microscillaceae Genus	+	+	+	-
NS9 marine group Genus			+	-	+
OLB12	+		+	-	+
Ohtaekwangia		+	+		+
Runella	-			-		+
Saprospiraceae Genus	-			-			+
Sediminibacterium	-
Sphingobacteriales Genus				-		+
Terrimonas				-		+	+
env OPS 17 Genus		-		-	-
Bdellovibrionota
Bacteriovoracaceae Genus	-			-		+
Bdellovibrio		-		-		+
OM27 clade				-		+	+
Oligoflexaceae Genus		-
Chloroflexi
Caldilineaceae Genus				-		+	+
JG30 KF CM45 Genus			+	-	+	+
KD4 96 Genus			+	-		+
RBG 13 54 9 Genus	-			-		+	+
SBR1031 Genus				-
Cyanobacteria
Candidatus Obscuribacter	-	-	+	-	+	+
Obscuribacteraceae Genus	+		+	-		+	-
Vampirovibrio			+	-	+	+
Vampirovibrionaceae Genus			+	-	+	+
Deinococcota
Deinococcus				-
Meiothermus				-
Dependentiae
Babeliaceae Genus	+			-	+	+
Desulfobacterota
Desulfobacterota Genus				-	+	+
Desulfobulbus				-			+
Elusimicrobiota
Lineage IIb Genus				-
Euryarchaeota
Methanobacteriaceae Genus				-	+
Fibrobacterota
Fibrobacteraceae Genus				-		+
Firmicutes
Acidaminobacter				-		+	+
Anaerospora			+
Bacillus		+	-	-
Brevibacillus		+
Clostridium sensu stricto 12				-		+
Clostridium sensu stricto 3			+	-	+	+
Enterococcus		+
Geothermomicrobium				-		+
Paraclostridium	+	+	+		-
Staphylococcus			+
Streptococcus				-		+
Thermoactinomycetaceae Genus	+		+
Vagococcus						+
ZOR0006			-		-	-
Fusobacteriota
Cetobacterium	+	+	-		-		-
Gemmatimonadota
Gemmatimonadaceae Genus		-		-	+	+
Halobacterota
Methanosarcina					+
MBNT15
MBNT15 Genus	+		+	-		+
Myxococcota
Aetherobacter			+	-	+
Anaeromyxobacter				-			+
Blfdi19 Genus				-
Haliangium			-	-		+	+
P3OB 42				-		+
Pajaroellobacter	+		+	-
Phaselicystis				-			+
Polyangia Genus				-
Sandaracinaceae Genus	+	+	+	-	+	+
mle1 27 Genus				-			+
NB1-j
NB1 j Genus		+	+	-	+	+
Nitrospirota
Nitrospira		-	+	-	+	+
Patescibacteria
Saccharimonadales Genus	+	+	+	-	+
TM7a					+
Planctomycetota
BD7 11 Genus				-			+
Blastopirellula	+			-	+	+
CL500 3			+	-	-	+
Fimbriiglobus	+	-	+	-	+	+
Gemmata		+	+	-	+	+
Gemmataceae Genus		-		-	+
Isosphaeraceae Genus			+	-	+	+	+
OM190 Genus				-	+	+	+
Pir4 lineage		-	+	-	+	+
Pirellula	+	+	+	-	-
Pirellulaceae Genus	+	-	+	-	-
Planctomycetales Genus			+	-	+	+
Planctomycetes Genus			+	-	+	+
Planctomycetota Genus				-		+
Rhodopirellula				-	+	+	-
SH PL14		+	+	-
SM1A02			+	-	+	+
Schlesneria		-		-	+	+
Telmatocola			+	-	+	+
Tundrisphaera	+	-	-	-			-
vadinHA49 Genus				-		+
Proteobacteria
A0839 Genus	+			-	+	+
AAP99				-		+
Acinetobacter	-
Aeromonas	-			+	+
Ahniella				-		+	+
Allorhizobium Neorhizobium Pararhizobium Rhizobium			-
Alphaproteobacteria Genus		+	+	-	+	+
Alsobacter		+	+	-	+	+
Aminobacter	-	-
Aquabacterium			-
Aquicella				-	+
Azospira				-	+	+
Bauldia	+		+	-	+	+
Beijerinckiaceae Genus		-	+	-		+	-
Blastomonas			-
Bosea	-	-	-	-
Burkholderia Caballeronia Paraburkholderia							+
Burkholderiales Genus				-
CCM19a Genus	+			-	+	+
Caedibacter	+		-		+
Candidatus Accumulibacter				-		+	-
Candidatus Alysiosphaera	+		+	-	+	+
Candidatus Berkiella		-	+		-
Candidatus Competibacter				-
Candidatus Megaira			+	-	+
Candidatus Ovatusbacter		-		-
Caulobacter			-
Chitinibacter		+	+		+		-
Chitinilyticum			-		-
Chitinivorax				-		+
Comamonadaceae Genus			-	-	+	+
Coxiella				-			+
Crenobacter		-
Curvibacter	-	-		-			+
Dechlorobacter				-		+
Dechloromonas				-		+
Defluviimonas		-		-	-
Diplorickettsiaceae Genus				-
Elstera	-					+
Fodinicurvataceae Genus				-		+
Ga0077536 Genus		-		-		+
Gammaproteobacteria Genus	-	-	+	-		+
Gemmobacter	+	+	-	-
Hirschia				-			+
Holosporaceae Genus	-	-			+
Hydrogenophaga							+
Hyphomicrobium		+	+
Ideonella				-		+	+
Inhella	-	-		-		+	+
Leeia	-	-		-		+
Legionella	-	-			-
Luteimonas	-			-		+	+
Magnetospirillum				-		+
Mesorhizobium				-
Methyloligellaceae Genus	+	+	+	-	+	+
Mycoplana	-	-	-
Neisseriaceae Genus			-
Nitrosomonadaceae Genus				-		+
Nitrosomonas		-		-		+
Nordella		-		-	+	+
Novosphingobium	-	-		-
Paucibacter	+	+	-	-	-
Pedomicrobium	+	+	+		-
Phenylobacterium				-			+
Phreatobacter				-	-
Plesiomonas	+	+			-	-	-
Polymorphobacter		-		-	+	+
Porticoccus				-		+
Pseudomonadales Genus	-	-	-	-	-	-
Pseudomonas		+			+
Pseudorhodobacter		-		-	+	+
Pseudorhodoplanes	+		+	-	-
Reyranella		-	+	-	+	+
Rheinheimera	-						+
Rhizobacter				-	+	+	+
Rhizobiaceae Genus	-	-		-
Rhizobiales Incertae Sedis Genus				-		+
Rhizorhapis	-			-			+
Rhodobacteraceae Genus				-	+	+
Rhodospirillales Genus				-			+
Rhodovarius				-	+	+
Rhodovastum		-		-
Rickettsiaceae Genus		+	+		+
Roseomonas	+	+	+	-	-	+
Rubrivivax				-	-
SC I 84 Genus		-		-	+	+
SWB02				-			+
Shewanella	-	-					-
Shinella	-	-		-
Sphingopyxis							+
Sphingorhabdus	-			-		+	+
Steroidobacter			+	-
Steroidobacteraceae Genus			+	-	+	+
Sulfurifustis							+
Sulfuritalea				-		+
Sutterellaceae Genus	+		-		-	-
TRA3 20 Genus							+
Tabrizicola			-	-	-		+
Undibacterium	-	-		-		+
Vibrio	-	-			+	+	-
Xanthobacteraceae Genus	-	-	+	-		+
Zoogloea				-		+	-
alphaI cluster	+		-		-
Spirochaetota
Turneriella	-	-		-		+
Verrucomicrobiota
Candidatus Protochlamydia				-	+	+
Chlamydiales Genus			+	-		+	-
Chthoniobacter		-		-	-	+	-
DEV008				-			+
Luteolibacter				-		+
Neochlamydia			+	-	-
Oikopleura			+	-	+	+
Opitutaceae Genus				-		+
Parachlamydiaceae Genus			-
Pedosphaeraceae Genus		-		-	+	+
SH3 11			-	-	-
Simkania		-
Simkaniaceae Genus			+
Verrucomicrobiaceae Genus	-	-		-			+
Verrucomicrobiae Genus			-
WPS-2
WPS 2 Genus	-						+

Session Info

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Session Info

## R version 4.3.3 (2024-02-29)
## Platform: x86_64-apple-darwin20 (64-bit)
## Running under: macOS 15.3
## 
## Matrix products: default
## BLAS:   /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vecLib.framework/Versions/A/libBLAS.dylib 
## LAPACK: /Library/Frameworks/R.framework/Versions/4.3-x86_64/Resources/lib/libRlapack.dylib;  LAPACK version 3.11.0
## 
## locale:
## [1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8
## 
## time zone: America/Los_Angeles
## tzcode source: internal
## 
## attached base packages:
## [1] grid      parallel  stats     graphics  grDevices utils     datasets 
## [8] methods   base     
## 
## other attached packages:
##  [1] lubridate_1.9.3       dplyr_1.1.4           readr_2.1.5          
##  [4] tidyr_1.3.1           tibble_3.2.1          tidyverse_2.0.0      
##  [7] multcomp_1.4-25       TH.data_1.1-2         MASS_7.3-60.0.1      
## [10] survival_3.6-4        mvtnorm_1.2-5         emmeans_1.10.1       
## [13] lme4_1.1-35.3         Matrix_1.6-5          car_3.1-2            
## [16] carData_3.0-5         broom_1.0.6           microViz_0.12.1      
## [19] picante_1.8.2         nlme_3.1-164          ape_5.8              
## [22] microbiome_1.24.0     phyloseqCompanion_1.1 vegan_2.6-4          
## [25] lattice_0.22-6        permute_0.9-7         stringr_1.5.1        
## [28] rlang_1.1.4           magrittr_2.0.3        GUniFrac_1.8         
## [31] doParallel_1.0.17     iterators_1.0.14      foreach_1.5.2        
## [34] data.table_1.15.4     phyloseq_1.46.0       Maaslin2_1.16.0      
## [37] ggnewscale_0.4.10     ComplexHeatmap_2.18.0 RColorBrewer_1.1-3   
## [40] gt_0.10.1             ggpubr_0.6.0          ggrepel_0.9.5        
## [43] ggExtra_0.10.1        ggbeeswarm_0.7.2      ggplot2_3.5.1        
## [46] furrr_0.3.1           future_1.33.2         purrr_1.0.2          
## [49] scales_1.3.0          rcompanion_2.4.35     knitr_1.45           
## [52] forcats_1.0.0        
## 
## loaded via a namespace (and not attached):
##   [1] fs_1.6.4                matrixStats_1.3.0       bitops_1.0-7           
##   [4] httr_1.4.7              insight_0.19.10         tools_4.3.3            
##   [7] backports_1.5.0         utf8_1.2.4              R6_2.5.1               
##  [10] mgcv_1.9-1              nortest_1.0-4           rhdf5filters_1.14.1    
##  [13] GetoptLong_1.0.5        withr_3.0.0             cli_3.6.2              
##  [16] Biobase_2.62.0          textshaping_0.3.7       logging_0.10-108       
##  [19] biglm_0.9-2.1           sandwich_3.1-0          labeling_0.4.3         
##  [22] sass_0.4.9              robustbase_0.99-2       pbapply_1.7-2          
##  [25] proxy_0.4-27            systemfonts_1.0.6       parallelly_1.37.1      
##  [28] readxl_1.4.3            rstudioapi_0.16.0       generics_0.1.3         
##  [31] shape_1.4.6.1           vroom_1.6.5             zip_2.3.1              
##  [34] inline_0.3.19           biomformat_1.30.0       fansi_1.0.6            
##  [37] DescTools_0.99.54       S4Vectors_0.40.2        abind_1.4-5            
##  [40] juicyjuice_0.1.0        lifecycle_1.0.4         yaml_2.3.8             
##  [43] rhdf5_2.46.1            Rtsne_0.17              promises_1.3.0         
##  [46] crayon_1.5.2            miniUI_0.1.1.1          cowplot_1.1.3          
##  [49] chromote_0.2.0          pillar_1.9.0            optparse_1.7.5         
##  [52] statip_0.2.3            rjson_0.2.21            boot_1.3-30            
##  [55] gld_2.6.6               estimability_1.5        codetools_0.2-20       
##  [58] glue_1.7.0              V8_4.4.2                vctrs_0.6.5            
##  [61] png_0.1-8               cellranger_1.1.0        gtable_0.3.5           
##  [64] datawizard_0.10.0       openxlsx_4.2.8          xfun_0.43              
##  [67] mime_0.12               libcoin_1.0-10          coda_0.19-4.1          
##  [70] rsconnect_1.3.4         pcaPP_2.0-4             modeest_2.4.0          
##  [73] timeDate_4032.109       pheatmap_1.0.12         statmod_1.5.0          
##  [76] bit64_4.0.5             modelbased_0.8.7        GenomeInfoDb_1.38.8    
##  [79] fBasics_4032.96         vipor_0.4.7             rpart_4.1.23           
##  [82] colorspace_2.1-0        BiocGenerics_0.48.1     DBI_1.2.3              
##  [85] ade4_1.7-22             processx_3.8.4          Exact_3.2              
##  [88] tidyselect_1.2.1        timeSeries_4032.109     curl_5.2.1             
##  [91] bit_4.0.5               compiler_4.3.3          expm_0.999-9           
##  [94] xml2_1.3.6              DEoptimR_1.1-3          lmtest_0.9-40          
##  [97] spatial_7.3-17          multcompView_0.1-10     digest_0.6.35          
## [100] minqa_1.2.6             rmarkdown_2.26          XVector_0.42.0         
## [103] htmltools_0.5.8.1       pkgconfig_2.0.3         openxlsx2_1.14         
## [106] stabledist_0.7-1        fastmap_1.2.0           GlobalOptions_0.1.2    
## [109] shiny_1.8.1.1           farver_2.1.2            zoo_1.8-12             
## [112] jsonlite_1.8.8          RCurl_1.98-1.14         modeltools_0.2-23      
## [115] GenomeInfoDbData_1.2.11 Rhdf5lib_1.24.2         munsell_0.5.1          
## [118] Rcpp_1.0.12             stringi_1.8.4           rootSolve_1.8.2.4      
## [121] stable_1.1.6            zlibbioc_1.48.2         plyr_1.8.9             
## [124] listenv_0.9.1           lmom_3.0                Biostrings_2.70.3      
## [127] splines_4.3.3           hash_2.2.6.3            multtest_2.58.0        
## [130] hms_1.1.3               circlize_0.4.16         ps_1.7.6               
## [133] igraph_2.0.3            ggsignif_0.6.4          reshape2_1.4.4         
## [136] stats4_4.3.3            rmutil_1.1.10           evaluate_0.23          
## [139] BiocManager_1.30.23     nloptr_2.0.3            tzdb_0.4.0             
## [142] httpuv_1.6.15           webshot2_0.1.1          getopt_1.20.4          
## [145] clue_0.3-65             coin_1.4-3              xtable_1.8-4           
## [148] e1071_1.7-14            rstatix_0.7.2           later_1.3.2            
## [151] class_7.3-22            ragg_1.3.0              report_0.5.8           
## [154] websocket_1.4.1         beeswarm_0.4.0          IRanges_2.36.0         
## [157] writexl_1.5.2           cluster_2.1.6           timechange_0.3.0       
## [160] globals_0.16.3